Mutant Trait Research Articles

Stochastic individual-based models with power law mutation rate on a general finite trait space

We consider a stochastic individual-based model for the evolution of a haploid, asexually reproducing population. The space of possible traits is given by the vertices of a (possibly directed) finite graph G=(V,E). The evolution of the population is driven by births, deaths, competition, and mutations along the edges of G. We are interested in the large population limit under a mutation rate μK given by a negative power of the carrying capacity K of the system: μK=K−1∕α,α>0. This results in several mutant traits being present at the same time and competing for invading the resident population. We describe the time evolution of the orders of magnitude of each sub-population on the logK time scale, as K tends to infinity. Using techniques developed in [8], we show that these are piecewise affine continuous functions, whose slopes are given by an algorithm describing the changes in the fitness landscape due to the succession of new resident or emergent types. This work generalises [25] to the stochastic setting, and Theorem 3.2 of [6] to any finite mutation graph. We illustrate our theorem by a series of examples describing surprising phenomena arising from the geometry of the graph and/or the rate of mutations.

Trait-dependent branching particle systems with competition and multiple offspring

In this work we model the dynamics of a population that evolves as a continuous time branching process with a trait structure and ecological interactions in form of mutations and competition between individuals. We generalize existing microscopic models by allowing individuals to have multiple offspring at a reproduction event. Furthermore, we allow the reproduction law to be influenced both by the trait type of the parent as well as by the mutant trait type. We look for tractable large population approximations. More precisely, under some natural assumption on the branching and mutation mechanisms, we establish a superprocess limit as solution of a well-posed martingale problem. Standard approaches do not apply in our case due to the lack of the branching property, which is a consequence of the dependency created by the competition between individuals. For showing uniqueness we therefore had to develop a generalization of Dawson's Girsanov Theorem that may be of independent interest.

Identification of wheat dwarf mutants and analysis on association between the mutant traits of the dwarf plants

<p indent=0mm>Dwarf mutant is an important gene resource in wheat breeding and plant height genetic research. In this paper, EMS (ethyl methyl sulfonate) was used to mutate the mature seeds of ‘Yunmai 53’, and 33 candidate dwarf mutants of M₃ generation were finally obtained by self-bred. Twenty-six dwarf mutants were selected by analyzing differences between the mutant parent in two years and that of M₂ and M₃ generation candidate plants, and the variation range of their plant height was from (13.61 ± <sc>0.11) cm</sc> to (44.08 ± <sc>1.73) cm.</sc> Twenty-six dwarf mutants with at least 2 mutant sites were verified based on 12 specific markers of 8 dwarf genes. In addition to plant height, 26 dwarf mutants also carried four mutational traits, spike length, spikelet density, internode number, and average internode length. The 26 dwarf mutants could be clustered into 5 subgroups. Among them, the first subgroup was the least in spikelets and florets, the second subgroup was the shortest in plant height, spike length and average internode length, and the highest spikelet density, while the third subgroup was the least in internode number. Plant height was significantly correlated with average internode length and internode number with partial correlation coefficients of 0.94 and 0.58, respectively, but not correlated with spike length, spikelet number and spikelet density. The plant height was genetically associated with internode length and internode number in 26 dwarf mutants. The mutants carried different combinations of the mutant gene, and can be useful in wheat dwarf breeding and the studies on genetic mechanism of the traits, such as plant height, spike length and spikelet density.

Genetic Parameters and Multivariate Analysis to Determine Secondary Traits in Selecting Wheat Mutant Adaptive on Tropical Lowlands

One of approaches to maintain the yield stability of the lowland tropical wheat is the use of secondary traits in the selection process. The identification of these characters requires a statistical approach in the form of genetic parameter analysis and multivariate analysis. The objective of this study was to determine the secondary traits of adaptive wheat mutants in the lowlands through the use of genetic parameters and multivariate analysis on the parameters. The study consisted of three field trials conducted in three different regencies, namely Jeneponto (135 m above sea level (asl)), Maros (100 m asl) and Bantaeng (125 m asl). The study used a nested design, where replications were nested in the environments. The genotype factors consisted of 20 genotypes repeated three times. 11 characters were observed including vegetative and reproductive characters. The analysis used consisted of repeatability, correlation, cluster analysis, principal component analysis, factor analysis, and cross print analysis. The overall results of the analysis indicate that the number of productive tillers is the main secondary trait for the selection of adaptive wheat in the lowlands. The character can be recommended for selection criteria in testing wheat lines in the lowlands to make an effective selection.

Genetic dissection of an allotetraploid interspecific CSSLs guides interspecific genetics and breeding in cotton

BackgroundThe low genetic diversity of Upland cotton limits the potential for genetic improvement. Making full use of the genetic resources of Sea-island cotton will facilitate genetic improvement of widely cultivated Upland cotton varieties. The chromosome segments substitution lines (CSSLs) provide an ideal strategy for mapping quantitative trait loci (QTL) in interspecific hybridization.ResultsIn this study, a CSSL population was developed by PCR-based markers assisted selection (MAS), derived from the crossing and backcrossing of Gossypium hirsutum (Gh) and G. barbadense (Gb), firstly. Then, by whole genome re-sequencing, 11,653,661 high-quality single nucleotide polymorphisms (SNPs) were identified which ultimately constructed 1211 recombination chromosome introgression segments from Gb. The sequencing-based physical map provided more accurate introgressions than the PCR-based markers. By exploiting CSSLs with mutant morphological traits, the genes responding for leaf shape and fuzz-less mutation in the Gb were identified. Based on a high-resolution recombination bin map to uncover genetic loci determining the phenotypic variance between Gh and Gb, 64 QTLs were identified for 14 agronomic traits with an interval length of 158 kb to 27 Mb. Surprisingly, multiple alleles of Gb showed extremely high value in enhancing cottonseed oil content (SOC).ConclusionsThis study provides guidance for studying interspecific inheritance, especially breeding researchers, for future studies using the traditional PCR-based molecular markers and high-throughput re-sequencing technology in the study of CSSLs. Available resources include candidate position for controlling cotton quality and quantitative traits, and excellent breeding materials. Collectively, our results provide insights into the genetic effects of Gb alleles on the Gh, and provide guidance for the utilization of Gb alleles in interspecific breeding.

Chloroplast development at low temperature requires the pseudouridine synthase gene TCD3 in rice

Low temperature affects a broad spectrum of cellular components in plants, such as chloroplasts, as well as plant metabolism. On the other hand, pseudouridine (Ψ) synthases are required for the most abundant post-transcriptional modification of RNA in Escherichia coli. However, the role of rice Ψ synthases in regulating chloroplast development at low temperature remains elusive. In this study, we identified the rice thermo-sensitive chlorophyll-deficient (tcd3) mutant, which displays an albino phenotype before the 4-leaf stage and ultimately dies when grown at 20 °C, but can grow normally at 32 °C. Genetic analysis showed that the mutant trait is controlled by a single recessive nuclear gene (tcd3). Map-based cloning, complementation and knockout tests revealed that TCD3 encodes a chloroplast-localized Ψ synthase. TCD3 is a cold-induced gene that is mainly expressed in leaves. The disruption of TCD3 severely affected the transcript levels of various chloroplast-associated genes, as well as ribosomal genes involved in chloroplast rRNA assembly at low temperature (20 °C), whereas the transcript levels of these genes were normal at high temperature (32 °C). These results provide a first glimpse into the importance of rice Ψ synthase gene in chloroplast development at low temperatures.

Optimal Virulence, Diffusion and Tradeoffs.

In this work we propose a variant of a classical SIR epidemiological model where pathogens are characterized by a (phenotypic) mutant trait x. Imposing that the trait x mutates according to a random walk process and that it directly influences the epidemiological components of the pathogen, we studied its evolutionary development by interpreting the tenet of maximizing the basic reproductive number of the pathogen as an optimal control problem. Pontryagin's maximum principle was used to identify the possible optimal evolutionary strategies of the pathogen. Qualitatively, three types of optimal evolutionary routes were identified and interpreted in the context of virulence evolution. Each optimal solution imposes a different tradeoff relation among the epidemiological parameters. The results predict (mostly) two kinds of infections: short-lasting mild infections and long-lasting acute infections.

Clinical Probe of Cyp2C8*2 Mutants in a Malaria Hyperendemic Zone: Evidence from North-Central, Nigeria.

A tremendous level of success has been achieved since the introduction of chloroquine and the combination of amodiaquine and artemisinin for the treatment of both complicated and uncomplicated malaria infections in sub-Saharan Africa. However, the recent discovery of drug resistant strains of Plasmodium falciparum (P.f.) and the ability of the parasite to ingest CYP2C8 into its digestive vacuole is of great public health concern. This study probes the occurrence of CYP2C8*2 allelic mutant amongst malaria patients in North-Central Nigeria. Three hundred and eighty five (385) unrelated study participants were screened for current malaria episodes using routine microscopy and/or rapid diagnostic test strips (RDTs). Chelex extraction method was used for single nucleotide polymorphisms (SNPs) and identification of CYP2C8*2 (805A > T) variant respectively. Wild-type (A) and the defective allele (T) were differentiated with the use of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP). The results obtained were further validated with Sanger sequencing of a few samples and thereafter, the genotype data were statistically processed. All alleles obtained were in Hardy Weinberg equilibrium. Out of the 385 participants (45.5% Male and 54.5% Female) genotyped for SNPs, 75 (19.5%) had the autosomal recessive mutant trait. Occurrence of mutant traits was gender and ethnic independent (p > 0.05). Yoruba ethnic group recorded a reduction in proportion of genotypic defective CYP2C8*2 allele (T) (1 in every 8 persons) with a carrier percentage of 13.3% compared with Hausa (26.62%); Igbo (25.37%) and other minority ethnic groups (17.6%). A remarkable inter-ethnic differences in autosomal recessive CYP2C8*2 allele was observed. By implication, there is a gradual incursion of genetic drift for poor CQ and AQ-Artemisinin metabolizers among the inhabitants.

Identification and Genetic Analysis of a Novel Allelic Variation of Brittle-1 with Endosperm Mutant in Maize

Endosperm mutants are critical to the studies on both starch synthesis and metabolism and genetic improvement of starch quality in maize. In the present study, a novel maize endosperm mutant A0178 of natural variation was used as the experimental material and identified and then characterized. Through phenotypic identification, genetic analysis, main ingredients measurement and embryo rescue, development of genetic mapping population from A0178, the endosperm mutant gene was located. The results showed that the mutant exhibited extremely low germination ability as attributed to the inhibited embryo development, and amounts of sugars were accumulated in the mutant seeds and more sugars content was detected at 23 days after pollination (DAP) in A0178 than B73. Employing genetic linkage analysis, the mutant trait was mapped in the bin 5.04 on chromosome 5. Sequence analysis showed that two sites of base transversion and insertion presented in the protein coding region and non-coding region of the mutant brittle-1 (bt1), the adenylate translocator encoding gene involved in the starch synthesis. The single base insertion in the coding region cause frameshift mutation, early termination and lose of function of Brittle-1 (BT1). All results suggested that bt1 is a novel allelic gene and the causal gene of this endosperm mutant, providing insights on the mechanism of endosperm formation in maize.

An efficient transgene-free DNA-editing system for Arabidopsis using a fluorescent marker.

To obtain transgene-free progeny by constructing a DNA editing system with a fluorescent screening marker gene and two pairs of single-guide RNAs to simultaneously recognize two different sites in the target gene encoding Arabidopsis microRNA(miR)160A RESULTS: The T1 seeds with red fluorescence were easily identified and were selected to verify that the proportion of miR160A knockout mutants reached approximately 50%. Seeds with no fluorescence in the T2 generation were selected and screened for homozygous mutants. In the T2 generation plants, the Cas9 fragment was not detected by polymerase chain reaction. The traits of the homozygous mutants were stably inherited by the T2 population. A highly efficient DNA-editing construct was successfully developed and can be used as a plant genome site-specific editing tool that may be useful for improving plant genetic resources.

Characterization and Genetic Mapping of White-Spotted Leaf (wspl) Mutant in Rice

Spotted leaf mutants which produce necrotic lesions spontaneously are important sources to study programmed cell death in plant defense responses. A novel white-spotted leaf (wspl) mutant was induced from Ilpum, Korean japonica rice cultivar by the treatment of ethyl methane sulfonate (EMS). The phenotype of wspl mutant differed from that of other spotted leaf mutants in that not only brown spots but also white lesion mimic spots were observed on the tip of the leaves from the vegetative stage. Strong nitro blue tetrazolium (NBT) and 3, 3′-diaminobenzidine (DAB) staining were observed on the older leaf of wspl mutant in microscopic reactive oxygen species (ROS) assay, and the chlorophyll content of wspl mutant maintained longer than wild-type in the old leaves. Genetic analysis revealed that the wspl mutant trait was controlled by a single recessive gene and the locus of wspl gene was mapped on the long arm of chromosome 5 between the flanking markers S05100 and S05112 (4.1 Mb). Through the combination of the genetic mapping and SNP analysis, two candidate genes for white-spotted leaf were identified in the genic region. A novel phenotype of white-spotted leaf mutant has not yet been reported, thus further study of the wspl mutant will contribute to understanding of the molecular mechanisms involved in lesion mimic phenotype in rice.

Mendelian inheritance of orange shell color in the Pacific oyster Crassostrea gigas

The oyster shell color has become an important factor influencing consumer choice. Understanding the genetic basis of shell colors is important in oyster genetic breeding. In this study, we investigated the inheritance of the orange shell color trait in the Pacific oyster. The oysters with orange shell color were generated in our breeding studies that has never been observed in natural population. It is a novel mutant trait in the Pacific oyster. In this study, we investigated the inheritance of the orange shell color trait crossing the orange variant with two other color variants (white and black). The shell color morph was determined for each in 27 first- and 12 s-generation crosses as well as 13 test crosses. We found that the orange shell color is recessive trait compared to black and white, and is determined by two independent recessive genes. In addition, our data revealed that the genetic loci controlling orange shell have no effects on the formation of black foreground pigment. The information obtained in this study could guide the breeding program for desired shell color in pacific oyster.

A SNP Mutation of SiCRC Regulates Seed Number Per Capsule and Capsule Length of cs1 Mutant in Sesame.

Seed number per capsule (SNC) is a major factor influencing seed yield and is an important trait with complex gene interaction effects. We first performed genetic analysis, gene cloning, and molecular mechanism study for an EMS-induced sesame mutant cs1 with fewer SNC and shorter capsule length (CL). The mutant traits were due to the pleiotropism of a regressive gene (Sics1). Capsule hormone determination showed that five out of 12 hormones, including auxin indole-3-acetic acid (IAA), had significantly different levels between wild type (WT) and mutant type (MT). KEGG pathway analysis showed that plant hormone signal transduction, especially the auxin signal transduction pathway, was the most abundant differentially expressed signaling pathway. After the cross-population association and regional genome screening, we found that three homozygous loci were retained in cs1. Further analysis of these three loci resulted in the identification of SiCRC as the candidate gene for cs1. SiCRC consists of seven exons and six introns encoding 163 amino acids. The SiCRC in cs1 showed a point mutation at intron 5 and exon 6 junction, resulting in the splice site being frame-shifted eight nucleotides further downstream, causing incorrect splicing. Taken together, we assumed the SNP mutation in SiCRC disrupted the function of the transcription factor, which might act downstream of the CRC-auxin signal transduction pathway, resulting in a shorter CL and less SNC mutation of cs1 in sesame. Our results highlight the molecular framework underlying the transcription factor CRC-mediated role of auxin transduction in SNC and CL development.

Selection of high-yielding, high-tech varieties of field pea (Pisum sativum L.)

One of the major challenges facing the selection of leguminous crops, and pea in particular, is to increase their technological properties and yield. An important problem is also the increase of protein content in the seeds. The aim of this work was to create a high-tech variety of field peas with a short growing period. By repeated individual selection from the hybrid population of the Chishminsky 95 × Usach field pea, a precocious, highly productive PamyatiHangildina variety of pea (leafless) was obtained. The combination of new mutant traits in one plant (seed shedding - def gene, leafless type - af gene) and elements of productivity structure with optimal level of their development allowed the variety to successfully pass all tests and in 2012 it was included in the State Register of Selection Achievements of the Russian Federation . For an average of 5 years of competitive testing, the variety had a seed yield of 1.62 t/ha, exceeding the standard (control) by 0.67 t/ha. The created variety has a short growing period and a high content of protein in the seeds. The research results could be successfully used in the selection of improved peas.

Selection of short stem Mentik Susu rice M3 from gamma ray irradiation

Mentik Susu is one of a local rice variety, which is not widely cultivated due to some unfavourable traits which were relatively high plant height and long harvest age. Efforts were done to overcome the problems such as planting mutation-induced plants through the utilization of gamma-ray irradiation. The objective of this research was to observe and select M3 mutants of Mentik Susu from gamma ray irradiation and obtain short-stemmed mutant plants. The research was conducted by simple design experiments (SDR). Established experimental plots for every irradiation doses and comparing the treatment result with the control to identify the effect of irradiation on the growth of Mentik Susu. The result showed that there were several strains with potential mutant traits based on the positive character of each individual. The M-MS2-15-3 strain with an irradiated dose of 200 grays has the shortest plant height of 92.10 cm with the highest productive tiller of 27.40 stems, has the highest seed yield per plant of 91.00 gram and has the shortest harvest age of 88 days. The M-MS2-15-4 strain with 200 gray irradiation dosehas the longest panicle length of 26.83 cm.

Identification and gene cloning of a male‐sterile mutant Oswbc11 in rice

AbstractCommercial application of nuclear genetic male sterility has been improved recently by a novel technique, Seed Production Technology (SPT), which incorporates transgenic maintainer lines capable of propagating non‐transgenic nuclear male‐sterile lines for use as female parents in hybrid production. Here, we identified a rice nuclear male‐sterile mutant, Oswbc11, with abnormal pollen development and lipid transport. We finely mapped the Oswbc11 gene into a 12.5‐kb region on chromosome 10 and found one candidate gene, which had a base substitution (C to T) resulting in a premature stop codon and was functionally confirmed by CRISPR/Cas9 technology. Gene OsWBC11 encodes a sub‐family member of adenosine triphosphate‐binding cassette transporter, which participates in the active transport of a wide range of molecules across membranes. Moreover, the agronomic traits of Oswbc11 mutants showed no significant differences compared to the wild‐type control except for the seed setting rate. These results indicated that Oswbc11 gene could be used in rice hybrid breeding as a recessive nuclear male‐sterile gene combined with CRISPR/Cas9 and the SPT technology and applied in different rice varieties.

Characterization of Non-heading Mutation in Heading Chinese Cabbage (Brassica rapa L. ssp. pekinensis).

Heading is a key agronomic trait of Chinese cabbage. A non-heading mutant with flat growth of heading leaves (fg-1) was isolated from an EMS-induced mutant population of the heading Chinese cabbage inbred line A03. In fg-1 mutant plants, the heading leaves are flat similar to rosette leaves. The epidermal cells on the adaxial surface of these leaves are significantly smaller, while those on the abaxial surface are much larger than in A03 plants. The segregation of the heading phenotype in the F2 and BC1 population suggests that the mutant trait is controlled by a pair of recessive alleles. Phytohormone analysis at the early heading stage showed significant decreases in IAA, ABA, JA and SA, with increases in methyl IAA and trans-Zeatin levels, suggesting they may coordinate leaf adaxial-abaxial polarity, development and morphology in fg-1. RNA-sequencing analysis at the early heading stage showed a decrease in expression levels of several auxin transport (BrAUX1, BrLAXs, and BrPINs) and responsive genes. Transcript levels of important ABA responsive genes, including BrABF3, were up-regulated in mid-leaf sections suggesting that both auxin and ABA signaling pathways play important roles in regulating leaf heading. In addition, a significant reduction in BrIAMT1 transcripts in fg-1 might contribute to leaf epinastic growth. The expression profiles of 19 genes with known roles in leaf polarity were significantly different in fg-1 leaves compared to wild type, suggesting that these genes might also regulate leaf heading in Chinese cabbage. In conclusion, leaf heading in Chinese cabbage is controlled through a complex network of hormone signaling and abaxial-adaxial patterning pathways. These findings increase our understanding of the molecular basis of head formation in Chinese cabbage.

Identification and gene mapping of a panicle apical abortion mutant (&amp;lt;italic&amp;gt;paa1331&amp;lt;/italic&amp;gt;) in rice

Rice panicle is an important agronomic organ which influences yield, and its development directly determines the yield potential. Over the past decades, a large number of genes related to grain size and grain number have been cloned, but there are few reports on cloning and function characterization of genes related to panicle development. We aim to explore the genes that regulate the degeneration of rice spike, improve the regulation network of panicle development, and provide theoretical basis for molecular breeding. In this study, a stably inherited mutant showing perturbed development of panicle ( panicle apical abortion 1331 ) was isolated from an ethyl methyl sulfate(EMS)mutagenized population of Yixiang 1B. Compared with the wild type, the mutant mainly showed a severe degeneration at the panicle tip (the affected florets account for as high as 53%), in addition to this, the plant height, tiller number and grain number were also affected. Observation on the development of panicle showed that the top spikelets of mutant paa1331 began to degrade during meiosis of the pollen mother cell, and the number of degenerated spikelets increased with the elongation of the young spike, distributing at the top of the first branch of the stem, while the number of degenerated spikelets showed a tendency to decrease gradually from top to bottom. Anatomical observations showed that the stamens of the mutant degenerated spikelets degenerated into linear, and the pulp was withered. The tissue section of mutant paa1331 and pollen staining assay showed that there were no pollen grains in the mutant anther, indicating that degeneration hindered the development of pollen; the vascular bundle of the mutant degenerated stem showed developmental defects, which may cause the transport of the material to be obstructed Trypan blue staining assay showed severe coloration of degraded spikelets on top of mutants, indicted that the panicle apical abortion was associated with programmed cell death. Hormone assay suggested that the IAA content of mutants was higher than that of wild type. Genetic analysis showed that mutant trait was controlled by a recessive gene. Combined the analysis of map-based cloning and De novo genome sequencing, we found one gene LOC_Os04g40720 , of which the fourth exon has an A-to-G single base substitution, which resulted in a change from Asn to Asp. At present, this gene has not been reported to confer function in panicle development, thus we treated it as candidate gene.

Rice SDSFL1 plays a critical role in the regulation of plant structure through the control of different phytohormones and altered cell structure

Semi-dwarfism is one of the most important agronomic traits for many cereal crops. In the present study, a mutant with semi-dwarf and short flag leaf 1, sdsfl1, was identified and characterized. The sdsfl1 mutant demonstrated some distinguished structural alterations, including shorter plant height and flag leaf length, increased tiller numbers and flag leaf width, and decreased panicle length compared with those of wild type (WT). Genetic analysis suggested that the mutant traits were completely controlled by a single recessive gene. The SDSFL1 gene was mapped to the long arm of chromosome 3 within a region of 44.6 kb between InDel markers A3P8.3 and A3P8.4. The DNA sequence analysis revealed that there was only a T to C substitution in the coding region of LOC_Os03g63970, resulting in the substitution of Tryptophan (Try) to Arginine (Arg) and encoding a GA 20 oxidase 1 protein of 372 amino acid residues. Photosynthesis analysis showed that the photosynthetic rate (Pn), stomatal conductance (Gs), and intercellular CO2 concentration (Ci) were significantly increased in sdsfl1. Chlorophyll a (Chl a), total Chl, and carotenoid contents were significantly increased in sdsfl1 compared with those in WT. sdsfl1 carried a reduced level of GA3 but reacted to exogenously applied gibberellins (GA). Moreover, the levels of abscisic acid (ABA), indole 3-acetic acid (IAA), and salicylic acid (SA) were notably improved in sdsfl1, whereas there was no noteworthy change in jasmonic acid (JA). The results thus offer a visible foundation for the molecular and physiological analysis of the SDSFL1 gene, which might participate in various functional pathways for controlling plant height and leaf length in rice breeding.

Detailed analysis of the prothoracic tissues transforming into wings in the Cephalothorax mutants of the Tribolium beetle

Despite the immense importance of the wing in the evolution and successful radiation of the insect lineages, the origin of this critical structure remains a hotly-debated mystery. Two possible tissues have been identified as an evolutionary origin of wings; the lateral expansion of the dorsal body wall (tergal edge) and structures related to an ancestral proximal leg segment (pleural tissues). Through studying wing-related tissues in the red flour beetle, Tribolium castaneum, we have previously presented evidence in support of a dual origin of insect wings, a third hypothesis proposing that wings evolved from a combination of both tergal and pleural tissues. One key finding came from the investigation of a Cephalothorax (Cx) mutant, in which the ectopic wing characteristic to this mutant was found to be formed from both tergal and pleural contributions. However, the degree of contribution of the two tissues to the wing remains elusive. Here, we took advantage of multiple Cx alleles available in Tribolium, and produced a variety of degrees and types of ectopic wing tissues in their prothoracic segments. Through detailed phenotypic scoring of the Cx phenotypes based on nine categories of mutant traits, along with comprehensive morphological analysis of the ectopic wing tissues, we found that (i) ectopic wing tissues can be formed at various locations in the prothorax, even internally, (ii) the lateral external ectopic wing tissues have tergal origin, while the internal and posterior external ectopic wing tissues appear to be of pleural origin, and (iii) the ectopic wing tissues of both tergal and pleural origin are capable of transforming into wing surface tissues. Collectively, these outcomes suggest that the evolutionary contribution of each tissue to a complete wing may be more complex than the simple binary view that is typically invoked by a dual origin model (i.e. the wing blade from the tergal contribution + musculature and articulation from the pleural contribution).