Reciprocal Hybrid Crosses Research Articles

A Scalable and Robust Chloroplast Genotyping Solution: Development and Application of SNP and InDel Markers in the Maize Chloroplast Genome.

Maize(Zea mays. L) is a globally important crop, and understanding its genetic diversity is crucial for plant breeding phylogenetic analyses and comparative genetics. While nuclear markers have been extensively used for mapping agriculturally important genes, they are limited in recognizing characteristics, such as cytoplasmic male sterility and reciprocal cross hybrids. In this study, we performed next-generation sequencing of 176samples, and the maize cultivars represented five distinct groups. A total of 89 single nucleotide polymorphisms (SNPs) and 11 insertion/deletion polymorphisms (InDels) were identified. To enable high-throughput detection, we successfully amplified and confirmed 49 SNP and InDel markers, which were defined as a Varietal Chloroplast Panel (VCP) using the Kompetitive Allele Specific PCR (KASP). The specific markers provided a valuable tool for identifying chloroplast groups. The verification experiment, focusing on the identification of reciprocal cross hybrids and cytoplasmic male sterility hybrids, demonstrated the significant advantages of VCP markers in maternal inheritance characterization. Furthermore, only a small subset of these markers is needed to provide useful information, showcasing the effectiveness of these markers in elucidating the artificial selection process of elite maize lines.

Temperature and sex shape reproductive barriers in a climate change hotspot.

Climate change is altering species ranges and reproductive interactions in existing ranges, offering species new scope to mate and hybridize. The outcomes will depend on how environmental factors shape reproductive barriers across life stages, yet this is rarely assessed across the environments that species encounter in nature. We assess prezygotic and postzygotic barriers, and their dependence on temperature and parental sex, in species of a reef-building tubeworm (Galeolaria) from a fast-warming biodiversity hotspot in southern Australia. By replicating pure and reciprocal hybrid crosses across 5 temperatures spanning species' thermal ranges, we estimate thermal tolerance curves (defining niches) for crosses and reproductive isolation at each temperature. By also replicating crosses at 3 life stages, we partition the contributions of prezygotic barriers at fertilization, postzygotic barriers at embryogenesis, and postzygotic barriers at larval development to reproductive isolation. We show that barriers are weaker at fertilization and embryogenesis, but stronger and more temperature sensitive at larval development, as species diverge in thermal niche. Asymmetry of barriers between parental sexes, moreover, suggests a complex interplay between niche differentiation and maternal inheritance. Our findings point to a key role for temperature in reproductive isolation, but also challenges for predicting the fate of isolation in future climates.

Unidirectional hybridization between American paddlefish Polyodon spathula (Walbaum, 1792) and sterlet Acipenser ruthenus (Linnaeus, 1758).

Interspecific hybridizations among sturgeon species are feasible and often bidirectional. The American paddlefish (Polyodon spathula) from Family Polyodontidae and sturgeon species from Family Acipenseridae were reported capable of hybridization, but viable hybrids have been described only in crosses with the American paddlefish as paternal parents. In the reciprocal cross, the hybrids were not viable however embryos start to develop and reach late gastrula and early neurula stages. The goal of this study was to examine the hybridization between the sterlet sturgeon (Acipenser ruthenus) and the American paddlefish. Hybrid and purebred crosses were produced by artificial fertilization. Viable hybrid offspring were harvested (three month old) and verified in the families produced by female sterlet crossing with male American paddlefish. In the reciprocal hybrid crosses with female American paddlefish and male sterlet, the embryos development did not pass over 120 h post fertilization, indicating the unidirectional hybridization between American paddlefish and sterlet. Chromosome counting showed for the same ploidy level of viable hybrid and parent species. Analysis of three microsatellite markers confirmed the unidirectional hybridization between the American paddlefish and the sterlet species. Overall, the inferred genetic cause suggests that unidirectional hybridization between American paddlefish and sterlet may be the case not only for these two species but likely also between American paddlefish and other sturgeon species.

Disproportionate role of nuclear-encoded proteins in organismal and mitochondrial thermal performance in a copepod.

Determining the mechanisms by which organisms evolve thermal tolerance is crucial to predicting how populations may respond to changes in local temperature regimes. Although evidence of relationships between mitochondrial background and thermal adaptation have been found, the presence of both nuclear-encoded and mitochondrial DNA (mtDNA)-encoded proteins warrants experiments aimed at parsing out the relative role of each genome in thermal adaptation. We investigated the relative role of mtDNA-encoded products in thermal tolerance between two divergent populations of Tigriopus californicus using first-generation (F1) hybrids that vary in maternally inherited mtDNA but are heterozygous for population-specific alleles across nuclear loci. We tested two measures of thermal tolerance, (1) survivorship to acute thermal stress and (2) thermal stability of mitochondrial performance in Complex I-fueled ATP synthesis, both across a range of increasing temperatures. We found that the southern population (San Diego, CA, USA) outperformed the northern population (Strawberry Hill, OR, USA) in survivorship, and that both reciprocal F1 hybrid crosses had intermediate survival. Mitochondria from the San Diego population displayed greater stability in ATP synthesis with increasing temperatures compared with those from Strawberry Hill. Interestingly, hybrids from both cross directions had synthesis profiles that were very similar to that of Strawberry Hill. Taken together, these results suggest that the relative role of the mtDNA in these phenotypes is negligible compared with that of elements encoded by nuclear DNA in this system.

Genetic control of growth and wood traits in Eucalyptus camaldulensis × E. urophylla and reciprocal hybrids

Eucalyptus camaldulensis (Ec) × E. urophylla (Eu) and reciprocal hybrid crosses were evaluated for growth and wood traits at four years at West Godavari Dt, Andhra Pradesh State located in Southern India, in relation to full and half sib progenies of pure E. camaldulensis maternal parents. Interspecific hybrid progenies had 16.6% higher average growth and 8% lower average wood basic density than pure species progeny. The interspecific hybrid progenies fibre properties viz., fibre length, fibre width, lumen diameter and fibre wall thickness were comparably higher than pure species progeny and check clones. The Ec × Eu crosses had comparable growth and wood basic density as the reciprocal hybrid progeny, but fibre length was 3.4% higher than the Eu × Ec hybrids, the values being statistically significant at p<.001 level. Analysis of variance of data was done using the 20th Edition of Genstat software (VSNi). Growth and fibre traits showed a positive relation but wood basic density showed a negative correlation (r = -0.94). There was a positive correspondence between the General Combining Ability (GCA) of pure E. camaldulensis maternal parents and their General Hybridizing Ability (GHA) for growth. The study showed that parents of Ec and Eu could be used to improve the growth and fibre traits in interspecific hybrids.

Tracking hybrid viability across life stages in a natural avian contact zone.

Hybrid inviability is an important post-zygotic reproductive barrier between species, but emerging signs of reduced viability can be difficult to study across the lifespan of natural hybrids. We use a combination of long-term monitoring, extra-pair paternity, and mitochondrial DNA identification in a natural hybrid zone of Ficedula flycatchers to detect emerging signs of intrinsic hybrid inviability across their entire lifespan. We evaluate possible evidence of Darwin's corollary to Haldane's rule, predicting asymmetries in inviability between hybrids resulting from reciprocal crosses, due to incompatible genetic factors with sex-specific inheritance patterns. We found higher hatching failure among mixed-species pairs, possibly indicating early developmental impairments associated with specific parental genetic combinations. Adult hybrids had a higher basal mortality rate than both parental species and different age-specific mortality trajectories. There were signs of differences in age-independent mortality rates between the reciprocal hybrid crosses: hybrids with a pied flycatcher mother experienced slightly increased mortality later in life. Using an exceptional dataset with many natural hybrids tracked across life stages, we provide evidence for several emerging signs of reduced hybrid viability. Incompatibilities between alleles located on autosomes and uniparentally inherited factors such as Z-linked and/or mitochondrial genes are strong candidates underlying intrinsic hybrid dysfunction in this system.

Heterosis for biomass yield and quality traits in a reciprocal cross population between Miscanthus sinensis and Miscanthus lutarioriparius

Heterosis plays an important role in plant breeding and has been widely used in the production of a variety of food crops. However, cross-breeding is particularly important in the breeding of new varieties of the energy crop miscanthus due to its characteristics of self-incompatibility and lack of interspecific reproductive isolation. Therefore, in the current study, 30 direct cross hybrids and 150 reciprocal cross hybrids were obtained from a cross between M. sinensis and M. lutarioriparius. These hybrids were significantly different in phenotype and were used for a detailed analysis of heterosis. The mid-parent, over-parent and over-standard heterosis of the hybrids for yield-related agronomic traits were analyzed. The results showed that the hybrids from direct and reciprocal crosses exhibited significant trait segregation and acquired dominant agronomic traits from both parents, showing mid-parent, over-parent and over-standard heterosis. Plant height, tiller number, stem node number, and stem diameter showed maternal inheritance. Principal component analysis was used to construct the linear combination relationship of yield-related agronomic traits. Excellent hybrids were screened against the commercial miscanthus variety Miscanthus × giganteus. These hybrids were superior to their two parents and M. × giganteus in terms of phenotypic yield-related agronomic traits, biomass yield, and quality. These results not only confirmed the feasibility of breeding miscanthus by remote crosses, but also provided a reference for the application, cross-breeding and breeding of new miscanthus varieties.

Allelic chromatin structure precedes imprinted expression of Kcnk9 during neurogenesis.

Differences in chromatin state inherited from the parental gametes influence the regulation of maternal and paternal alleles in offspring. This phenomenon, known as genomic imprinting, results in genes preferentially transcribed from one parental allele. While local epigenetic factors such as DNA methylation are known to be important for the establishment of imprinted gene expression, less is known about the mechanisms by which differentially methylated regions (DMRs) lead to differences in allelic expression across broad stretches of chromatin. Allele-specific higher-order chromatin structure has been observed at multiple imprinted loci, consistent with the observation of allelic binding of the chromatin-organizing factor CTCF at multiple DMRs. However, whether allelic chromatin structure impacts allelic gene expression is not known for most imprinted loci. Here we characterize the mechanisms underlying brain-specific imprinted expression of the Peg13-Kcnk9 locus, an imprinted region associated with intellectual disability. We performed region capture Hi-C on mouse brains from reciprocal hybrid crosses and found imprinted higher-order chromatin structure caused by the allelic binding of CTCF to the Peg13 DMR. Using an in vitro neuron differentiation system, we showed that imprinted chromatin structure precedes imprinted expression at the locus. Additionally, activation of a distal enhancer induced imprinted expression of Kcnk9 in an allelic chromatin structure-dependent manner. This work provides a high-resolution map of imprinted chromatin structure and demonstrates that chromatin state established in early development can promote imprinted expression upon differentiation.

Sex-specific estimation of cis and trans regulation of gene expression in heads and gonads of Drosophila melanogaster.

The regulatory architecture of gene expression is known to differ substantially between sexes in Drosophila, but most studies performed so far used whole-body data and only single crosses, which may have limited their scope to detect patterns that are robust across tissues and biological replicates. Here, we use allele-specific gene expression of parental and reciprocal hybrid crosses between 6 Drosophila melanogaster inbred lines to quantify cis- and trans-regulatory variation in heads and gonads of both sexes separately across 3 replicate crosses. Our results suggest that female and male heads, as well as ovaries, have a similar regulatory architecture. On the other hand, testes display more and substantially different cis-regulatory effects, suggesting that sex differences in the regulatory architecture that have been previously observed may largely derive from testis-specific effects. We also examine the difference in cis-regulatory variation of genes across different levels of sex bias in gonads and heads. Consistent with the idea that intersex correlations constrain expression and can lead to sexual antagonism, we find more cis variation in unbiased and moderately biased genes in heads. In ovaries, reduced cis variation is observed for male-biased genes, suggesting that cis variants acting on these genes in males do not lead to changes in ovary expression. Finally, we examine the dominance patterns of gene expression and find that sex- and tissue-specific patterns of inheritance as well as trans-regulatory variation are highly variable across biological crosses, although these were performed in highly controlled experimental conditions. This highlights the importance of using various genetic backgrounds to infer generalizable patterns.

Asymmetric gene expression in grain development of reciprocal crosses between tetraploid and hexaploid wheats

Production of viable progeny from interploid crosses requires precise regulation of gene expression from maternal and paternal chromosomes, yet the transcripts contributed to hybrid seeds from polyploid parent species have rarely been explored. To investigate the genome-wide maternal and paternal contributions to polyploid grain development, we analyzed the transcriptomes of developing embryos, from zygote to maturity, alongside endosperm in two stages of development, using reciprocal crosses between tetraploid and hexaploid wheats. Reciprocal crosses between species with varied levels of ploidy displayed broad impacts on gene expression, including shifts in alternative splicing events in select crosses, as illustrated by active splicing events, enhanced protein synthesis and chromatin remodeling. Homoeologous gene expression was repressed on the univalent D genome in pentaploids, but this suppression was attenuated in crosses with a higher ploidy maternal parent. Imprinted genes were identified in endosperm and early embryo tissues, supporting predominant maternal effects on early embryogenesis. By systematically investigating the complex transcriptional networks in reciprocal-cross hybrids, this study presents a framework for understanding the genomic incompatibility and transcriptome shock that results from interspecific hybridization and uncovers the transcriptional impacts on hybrid seeds created from agriculturally-relevant polyploid species.

Co-expression network analysis of environmental canalization in the ascidian Ciona

BackgroundCanalization, or buffering, is defined as developmental stability in the face of genetic and/or environmental perturbations. Understanding how canalization works is important in predicting how species survive environmental change, as well as deciphering how development can be altered in the evolutionary process. However, how developmental gene expression is linked to buffering remains unclear. We addressed this by co-expression network analysis, comparing gene expression changes caused by heat stress during development at a whole-embryonic scale in reciprocal hybrid crosses of sibling species of the ascidian Ciona that are adapted to different thermal environments.ResultsSince our previous work showed that developmental buffering in this group is maternally inherited, we first identified maternal developmental buffering genes (MDBGs) in which the expression level in embryos is both correlated to the level of environmental canalization and also differentially expressed depending on the species’ gender roles in hybrid crosses. We found only 15 MDBGs, all of which showed high correlation coefficient values for expression with a large number of other genes, and 14 of these belonged to a single co-expression module. We then calculated correlation coefficients of expression between MDBGs and transcription factors in the central nervous system (CNS) developmental gene network that had previously been identified experimentally. We found that, compared to the correlation coefficients between MDBGs, which had an average of 0.96, the MDBGs are loosely linked to the CNS developmental genes (average correlation coefficient 0.45). Further, we investigated the correlation of each developmental to MDBGs, showing that only four out of 62 CNS developmental genes showed correlation coefficient > 0.9, comparable to the values between MDBGs, and three of these four genes were signaling molecules: BMP2/4, Wnt7, and Delta-like.ConclusionsWe show that the developmental pathway is not centrally located within the buffering network. We found that out of 62 genes in the developmental gene network, only four genes showed correlation coefficients as high as between MDBGs. We propose that loose links to MDBGs stabilize spatiotemporally dynamic development.

Study on the Effect of Mass Selection and Hybridization on Growth Performance of Chinese Pearl Oyster Pinctada martensii

The pearl oyster Pinctada martensii is an important species for sea pearl production in China. To explore the influence of the combination of mass selection and hybridization on growth performance of P. martensii, we established four selected groups and four control groups (each with two within-family crosses and two reciprocal hybrid crosses) using 1-year-old offspring of two families (Family A and Family B) from Beihai, Guangxi Province, China. Generally, the growth of the selected group was greater than that of the control group for both within-family crosses and reciprocal hybrid crosses. Shell length and width were affected by genotype, environmental factors, and the interaction between the two on Days 210 and 360. The shell widths of the four reciprocal hybrid crosses all showed heterosis on Day 360. The four within-family crosses showed a certain degree of inbreeding depression during the growth period. On Day 360, the three genetic parameters for shell width of the selected group of ♀ Family B and ♂ Family A were the largest, with values of 0.70, 1.17, and 0.06 for realized heritability, standard response to selection, and current genetic gain, respectively. Overall, the growth performance of ♀ Family B × ♂ Family A in the selected group was the best. Therefore, the combination of mass selection and hybridization could be an effective way to improve the growth performance of P. martensii.

Mitonuclear interactions alter sex-specific longevity in a species without sex chromosomes.

Impaired mitochondrial function can lead to senescence and the ageing phenotype. Theory predicts degenerative ageing phenotypes and mitochondrial pathologies may occur more frequently in males due to the matrilineal inheritance pattern of mitochondrial DNA observed in most eukaryotes. Here, we estimated the sex-specific longevity for parental and reciprocal F1 hybrid crosses for inbred lines derived from two allopatric Tigriopus californicus populations with over 20% mitochondrial DNA divergence. T. californicus lacks sex chromosomes allowing for more direct testing of mitochondrial function in sex-specific ageing. To better understand the ageing mechanism, we estimated two age-related phenotypes (mtDNA content and 8-hydroxy-20-deoxyguanosine (8-OH-dG) DNA damage) at two time points in the lifespan. Sex differences in lifespan depended on the mitochondrial and nuclear backgrounds, including differences between reciprocal F1 crosses which have different mitochondrial haplotypes on a 50 : 50 nuclear background, with nuclear contributions coming from alternative parents. Young females showed the highest mtDNA content which decreased with age, while DNA damage in males increased with age and exceed that of females 56 days after hatching. The adult sex ratio was male-biased and was attributed to complex mitonuclear interactions. Results thus demonstrate that sex differences in ageing depend on mitonuclear interactions in the absence of sex chromosomes.

The Effect of Hybridization between Natural and Cultivated Peruvian Scallop Argopecten purpuratus populations on Growth and Tolerance to Abiotic Stress

Intraspecific hybridization has been a strategy used frequently in mollusc aquaculture to improve the performance of traits for productive interest. In this study, the effect of hybridization between a natural population (Arica = A) and a cultivated population (Coquimbo = C; without genetic management for more than 30 y) of the Peruvian scallop Argopecten purpuratus was evaluated to assess the effect of this strategy on the performance of the cultivated population in terms of growth, survival, and stress tolerance due to hypoxia and increased temperature. To this end, progenies from intrapopulation (C × C and A × A) and reciprocal hybrid (C × A and A × C) crosses were produced and were evaluated after 6 mo under sea cultivation and in a controlled environment. Stress responses were evaluated via metabolic rates and the transcriptional level of stress-associated genes [superoxide dismutase (SOD) and HSP70]. The results showed that CC and the reciprocal hybrids CA and AC had greater size and survival rates than AA; on the other hand, AA presented the highest metabolic rates when exposed to stress; and, in parallel, AA and both hybrids (CA and AC) presented greater antioxidant capacity (SOD) when exposed to hyperthermia. Therefore, growth and survival showed positive heterosis because the hybrids performed better in these traits than the parental mean. In addition, hybridization with the Arica population would allow the Coquimbo stock under cultivation to improve its resilience to temperature increases associated to oceanographic oscillations or to climate change.

Maternal effects shape the alternative splicing of parental alleles in reciprocal cross hybrids of Megalobrama amblycephala \xd7 Culter alburnus

BackgroundMaternal effects contribute to adaptive significance for shaping various phenotypes of many traits. Potential implications of maternal effects are the cause of expression diversity, but these effects on mRNA expression and alternative splicing (AS) have not been fully elucidated in hybrid animals.ResultsTwo reciprocal cross hybrids following hybridization of Megalobrama amblycephala (blunt snout bream, BSB) and Culter alburnus (topmouth culter, TC) were used as a model to investigate maternal effects. By comparing the expression of BSB- and TC- homoeologous genes between the two reciprocal cross hybrids, we identified 49–348 differentially expressed BSB-homoeologous genes and 54–354 differentially expressed TC-homoeologous genes. 2402, 2959, and 3418 AS events between the two reciprocal cross hybrids were detected in Illumina data of muscle, liver, and gonad, respectively. Moreover, 21,577 (TC-homoeologs) and 30,007 (BSB-homoeologs) AS events were found in the 20,131 homoeologous gene pairs of TBF3 based on PacBio data, while 30,561 (TC-homoeologs) and 30,305 (BSB-homoeologs) AS events were found in BTF3. These results further improve AS prediction at the homoeolog level. The various AS patterns in bmpr2a belonging to the bone morphogenetic protein family were selected as AS models to investigate the expression diversity and its potential effects to body shape traits.ConclusionsThe distribution of differentially expressed genes and AS in BSB- and TC-subgenomes exhibited various changes between the two reciprocal cross hybrids, suggesting that maternal effects were the cause of expression diversity. These findings provide a novel insight into mRNA expression changes and AS under maternal effects in lower vertebrates.

Are you more than the sum of your parents' genes? Phenotypic plasticity in a clonal vertebrate and F1 hybrids of its parental species.

All known vertebrate clones have originated from hybridization events and some have produced distinct evolutionary lineages via hybrid speciation. Amazon mollies (Poecilia formosa) present an excellent study system to investigate how clonal species have adapted to heterogeneous environments because they are the product of a single hybridization event between male sailfin mollies (Poecilia latipinna) and female Atlantic mollies (Poecilia mexicana). Here, we ask whether the hybrid species differs from the combination of its parental species' genes in its plastic response to different environments. Using a three-way factorial design, we exposed neonates produced by Amazon mollies and reciprocal F1 hybrid crosses to different thermal (24°C and 29°C) and salinity (0/2, 12, and 20 ppt) regimes. We measured various ontogenetic and life history characteristics across the life span of females. Our major results were as follows: (1) Reaction norms of growth and maturation to temperature and salinity are quite similar between the two hybrid crosses; (2) Amazon molly reaction norms were qualitatively different than the P. latipinna male and P. mexicana female (L×M) hybrids for the ontogenetic variables; (3) Amazon molly reaction norms in reproductive traits were also quite different from L×M hybrids; and (4) The reaction norms of net fertility were very different between Amazon mollies and L×M hybrids. We conclude that best locale for Amazon mollies is not the best locale for hybrids, which suggests that Amazon mollies are not just an unmodified mix of parental genes but instead have adapted to the variable environments in which they are found. Hybridization resulting in asexuality may represent an underappreciated mechanism of speciation because the unlikely events required to produce such hybrids rarely occur and is dependent upon the genetic distance between parental species.

Allele-specific expression and alternative splicing in horse×donkey and cattle×yak hybrids.

Divergence of gene expression and alternative splicing is a crucial driving force in the evolution of species; to date, however the molecular mechanism remains unclear. Hybrids of closely related species provide a suitable model to analyze allele-specific expression (ASE) and allele-specific alternative splicing (ASS). Analysis of ASE and ASS can uncover the differences in cis-regulatory elements between closely related species, while eliminating interference of trans-regulatory elements. Here, we provide a detailed characterization of ASE and ASS from 19 and 10 transcriptome datasets across five tissues from reciprocal-cross hybrids of horse×donkey (mule/hinny) and cattle×yak (dzo), respectively. Results showed that 4.8%–8.7% and 10.8%–16.7% of genes exhibited ASE and ASS, respectively. Notably, lncRNAs and pseudogenes were more likely to show ASE than protein-coding genes. In addition, genes showing ASE and ASS in mule/hinny were found to be involved in the regulation of muscle strength, whereas those of dzo were involved in high-altitude adaptation. In conclusion, our study demonstrated that exploration of genes showing ASE and ASS in hybrids of closely related species is feasible for species evolution research.

Behavioural mechanisms of sexual isolation involving multiple modalities and their inheritance.

Speciation research dissects the genetics and evolution of reproductive barriers between parental species. Hybrids are the "gatekeepers" of gene flow, so it is also important to understand the behavioural mechanisms and genetics of any potential isolation from their parental species. We tested the role of multiple behavioural barriers in reproductive isolation among closely related field crickets and their hybrids (Teleogryllus oceanicus and Teleogryllus commodus). These species hybridize in the laboratory, but the behaviour of hybrids is unusual and there is little evidence for gene flow in the wild. We found that heterospecific pairs exhibited reduced rates of courtship behaviour due to discrimination by both sexes, and that this behavioural isolation was symmetrical. However, hybrids were not sexually selected against and exhibited high rates of courtship behaviour even though hybrid females are sterile. Using reciprocal hybrid crosses, we characterized patterns of interspecific divergence and inheritance in key sexual traits that might underlie the mating patterns we found: calling song, courtship song and cuticular hydrocarbons (CHCs). Song traits exhibited both sex linkage and transgressive segregation, whereas CHCs exhibited only the latter. Calculations of the strength of isolation exerted by these sexual traits suggest that close-range signals are as important as long-distance signals in contributing to interspecific sexual isolation. The surprisingly weak mating barriers observed between hybrids and parental species highlight the need to examine reproductive isolating mechanisms and their genetic bases across different potential stages of introgressive hybridization.

Morphological and Molecular Analyses of Reciprocal Hybrids between ‘Slim Whitman’ and ‘Pinza’, Two Narcissus pseudonarcissus Cultivars

Narcissus pseudonarcissus, also known as daffodil, is a world-famous ornamental flower. In this study, for the first time, cross-pollinations between two widely cultivated N. pseudonarcissus varieties ‘Slim Whitman’ and ‘Pinza’ were performed. After eight consecutive years of cultivation, 27 reciprocal hybrids with different genotypes survived; 15 hybrids in ‘Slim Whitman’ × ‘Pinza’ and 12 in ‘Pinza’ × ‘Slim Whitman’. Twenty ornamental and agronomic characters were observed to evaluate the pattern and extent of genetic variability of the hybrids and relatedness with their parents. The hybrids showed great variation in most morphological characters compared with the parents, especially in leaf and flower characters. Hybrids SP03, SP04, SP05, SP12, SP13, PS04, PS06, PS07, PS08, and PS11 had evident growth advantage in some aspects compared with both parents. Of these hybrids, SP04 got novel flowers with white petals and an yellow-orange corona, and had the potential to become a new popular N. pseudonarcissus cultivar. Hybrids SP01, SP03, SP05, PS04, PS06, and PS07 also possessed a great ornamental value. Using cluster analysis based on morphological traits and random amplified polymorphic DNA (RAPD) molecular markers, genetic relationships among the reciprocal cross hybrids and their parents were further analyzed. The 27 reciprocal hybrids and their parents grouped into divergent clusters, showing that there was rich genetic variation among the hybrids tested. This study will pave the way for hybridization breeding programs of N. pseudonarcissus.

Inheritance of Monocrotophos Resistance in Egg Parasitoid Trichogramma Chilonis (Ishii) (Hymenoptera:Trichogrammatidae)

Inheritance of monocrotophos resistance was studied in the laboratory population of T. chilonis. After 10 cycles of selection pressure under laboratory condition, the resistant strain acquired LC50 value of 0.346mL compared toLC50 value of 0.114 mL in the susceptible strain. Evidence from bioassay of F1 reciprocal hybrid crosses backcrossed with respective resistant 'R' and susceptible 'S'parentalstrains of T. chilonis, aimed to determine mode of inheritance of insecticide tolerance indicated that F1 crosses exhibited a complete dominant response to monocrotophos, with degree of dominance value (D) of 1.62. The resistance factor (Rf) of resistant strain was 3.04 folds and of F1 crosses were 3.675 folds over susceptible strain. Result of this study suggests resistance to the insecticide was probably controlled by a single gene. These results provide the basic information for designing successful management programmes for the control of Lepidopteran pests using resistant strain as a component of IPM.