Interspecific Hybrids Research Articles

Utilisation of GMS lines for production ofheterotic bacterial wilt resistant hybrids in <i>Tagetes</i> spp.

The study was conducted using six testers, of which two (KAU M-1 and KAU M-2) were completely bacterial wilt resistant, and four genetic male sterile lines (GMS), in a line x tester model and further the F1 hybrids were evaluated in a bacterial wilt sick plot for various vegetative, floral and yield parameters along with their response to wilt incidence. Field evaluation for bacterial wilt resistance revealed that out of 24 hybrids, 13 hybrids involving KAU M-1 and KAU M-2 as testers exhibited bacterial wilt resistance. Analysis on heterosis for various characters viz., vegetative, floral, yield and total carotenoids content showed significant heterosis for many of the hybrids over mid parent, better parent and the check variety. The highest standard heterosis for yield was observed for the interspecific hybrid KU Ms-6 xx KAU M-47 (190.84%) followed by an intraspecific hybrid KAU Ms-42 x KAU M-2 (70.93%). Based on the wilt resistance and heterotic performance, these two hybrids can be suggested for commercial cultivation in wilt affected areas.

Genetic relationships of Michelia compressa (Magnoliaceae) with Michelia species and its improvement by interspecific hybridization

Genetic relationships of Michelia compressa (Magnoliaceae) with Michelia species and its improvement by interspecific hybridization

Towards Reliable Detection of Introgression in the Presence of Among-Species Rate Variation.

The role of interspecific hybridization has recently seen increasing attention, especially in the context of diversification dynamics. Genomic research has now made it abundantly clear that both hybridization and introgression-the exchange of genetic material through hybridization and backcrossing-are far more common than previously thought. Besides cases of ongoing or recent genetic exchange between taxa, an increasing number of studies report "ancient introgression"- referring to results of hybridization that took place in the distant past. However, it is not clear whether commonly used methods for the detection of introgression are applicable to such old systems, given that most of these methods were originally developed for analyses at the level of populations and recently diverged species, affected by recent or ongoing genetic exchange. In particular, the assumption of constant evolutionary rates, which is implicit in many commonly used approaches, is more likely to be violated as evolutionary divergence increases. To test the limitations of introgression detection methods when being applied to old systems, we simulated thousands of genomic datasets under a wide range of settings, with varying degrees of among-species rate variation and introgression. Using these simulated datasets, we showed that some commonly applied statistical methods, including the D-statistic and certain tests based on sets of local phylogenetic trees, can produce false-positive signals of introgression between divergent taxa that have different rates of evolution. These misleading signals are caused by the presence of homoplasies occurring at different rates in different lineages. To distinguish between the patterns caused by rate variation and genuine introgression, we developed a new test that is based on the expected clustering of introgressed sites along the genome and implemented this test in the program Dsuite.

Selection of Saccharum spp. × Saccharum spontaneum progeny for sugar and biomass traits

ABSTRACT This study evaluated an interspecific hybrid (Saccharum spp. × S. spontaneum) population with the aim of developing germplasm for use in both sugar and bioenergy (biomass) production. Specific objectives were to determine the genetic variability and heritability among the interspecific hybrid progenies for traits of economic importance and to use the Best Linear Unbiased Predictions (BLUPs) to determine their genotypic values. The 46 interspecific hybrids used in this study originated from crossing commercial sugarcane cultivars (UT5, F152, and KK07–559) as female and S. spontaneum clones (THS98–41, THS98–91, THS98–95, THS98–94, and THS97–51) as male parents. There were significant residual variances, attributed to the small plot size, but the juice quality traits were measured with relatively higher precision than other traits. Selection recommendations varied for different traits, suggesting selection pressure could be moderate to high for juice quality traits and moderate for yield-related traits. No hybrid genotypes were found to outperform cultivar parents for traits such as commercial cane sugar and low fiber, necessitating further germplasm enhancement for improvement of varieties destined for sugar production. On the contrary, transgressive segregation was observed for cane yield and other traits influencing biomass yield. The production potential of these hybrids could be tested in larger plots across multiple locations.

Expansion of learning capacity elicited by interspecific hybridization.

Learned behavior, a fundamental adaptive trait in fluctuating environments, is shaped by species-specific constraints. This phenomenon is evident in songbirds, which acquire their species-specific songs through vocal learning. To explore the neurogenetic mechanisms underlying species-specific song learning, we generated F1 hybrid songbirds by crossing Taeniopygia guttata with Aidemosyne modesta. These F1 hybrids demonstrate expanded learning capacities, adeptly mimicking songs from both parental species and other heterospecific songs more extensively than their parental counterparts. Despite the conserved size of brain regions and neuron numbers in the neural circuits for song learning and production, single-cell transcriptomics reveals distinctive transcriptional characteristics in the F1 hybrids, especially in vocal-motor projection neurons. These neurons exhibit enrichment for nonadditively expressed genes, particularly those related to ion channel activity and cell adhesion, which are associated with the degree of song learning among F1 individuals. Our findings provide insights into the emergence of altered learning capabilities through hybridization, linked to cell type-specific transcriptional changes.

Exploring genetic variability and molecular-cytogenetic profiling of rye-wheat derivatives

Interspecific plant hybridisation is a pivotal evolutionary process shaping speciation dynamics. In the context of bread wheat cultivation in India, stripe rust stands out as the predominant factor contributing to yield reduction. With the objective of improving stripe rust resistance and floral traits, diverse back-crossed population was meticulously developed through triticale-wheat crosses. The BC2F7 generation of rye-wheat derivatives underwent a comprehensive analysis encompassing genetic variability, trait associations, multiple regressions, and principal component, with a focus on floral and yield attributes. The meticulously chosen stripe rust-resistant derivative lines underwent comprehensive evaluation, including an assessment of chromosomal abnormalities, fluorescence in situ hybridisation, and rye genome-specific molecular markers analysis. The statistical analysis substantiates that the derived population exhibits significant variations for all the examined traits. Positive correlations were observed with anther size, pollen viability, tiller number, thousand kernel weight, and grain yield. Multiple regression analysis reveals that anther size, pollen viability, tiller number, and thousand kernel weights emerge as pivotal contributors influencing grain yield. Moreover, the three principal components exhibit more than one eigen-value, thereby signifying their capacity to encapsulate the maximum variability within the dataset. In addition, chromosomal abnormalities, encompassing univalent, multivalent, laggards, bridge chromosomes, and micronuclei, were observed within the PMCs of rye wheat derivative lines. Fluorescence in-situ hybridisation unequivocally demonstrated rye introgression by green signals. Furthermore, the affirmation of rye introgression was corroborated by rye genome-specific molecular markers. Hence, Comprehensive methods unveil promising traits in rye-wheat lines, advancing wheat breeding in India.

Multi-omic analysis reveals the effects of interspecific hybridization on the synthesis of seed reserve polymers in a Triticum turgidum ssp. durum × Aegilops sharonensis amphidiploid

BackgroundWheat grain endosperm is mainly composed of proteins and starch. The contents and the overall composition of seed storage proteins (SSP) markedly affect the processing quality of wheat flour. Polyploidization results in duplicated chromosomes, and the genomes are often unstable and may result in a large number of gene losses and gene rearrangements. However, the instability of the genome itself, as well as the large number of duplicated genes generated during polyploidy, is an important driving force for genetic innovation. In this study, we compared the differences in starch and SSP, and analyzed the transcriptome and metabolome among Aegilops sharonensis (R7), durum wheat (Z636) and amphidiploid (Z636×R7) to reveal the effects of polyploidization on the synthesis of seed reserve polymers.ResultsThe total starch and amylose content of Z636×R7 was significantly higher than R7 and lower than Z636. The gliadin and glutenin contents of Z636×R7 were higher than those in Z636 and R7. Through transcriptome analysis, there were 21,037, 2197, 15,090 differentially expressed genes (DEGs) in the three comparison groups of R7 vs Z636, Z636 vs Z636×R7, and Z636×R7 vs R7, respectively, which were mainly enriched in carbon metabolism and amino acid biosynthesis pathways. Transcriptome data and qRT-PCR were combined to analyze the expression levels of genes related to storage polymers. It was found that the expression levels of some starch synthase genes, namely AGP-L, AGP-S and GBSSI in Z636×R7 were higher than in R7 and among the 17 DEGs related to storage proteins, the expression levels of 14 genes in R7 were lower than those in Z636 and Z636×R7. According to the classification analysis of all differential metabolites, most belonged to carboxylic acids and derivatives, and fatty acyls were enriched in the biosynthesis of unsaturated fatty acids, niacin and nicotinamide metabolism, one-carbon pool by folate, etc.ConclusionAfter allopolyploidization, the expression of genes related to starch synthesis was down-regulated in Z636×R7, and the process of starch synthesis was inhibited, resulting in delayed starch accumulation and prolongation of the seed development process. Therefore, at the same development time point, the starch accumulation of Z636×R7 lagged behind that of Z636. In this study, the expression of the GSe2 gene in Z636×R7 was higher than that of the two parents, which was beneficial to protein synthesis, and increased the protein content. These results eventually led to changes in the synthesis of seed reserve polymers. The current study provided a basis for a greater in-depth understanding of the mechanism of wheat allopolyploid formation and its stable preservation, and also promoted the effective exploitation of high-value alleles.

Microscopic observation and transcriptome analysis provide insights into mechanisms of hybrid incompatibility in Rhododendron

Hybridization across species has been widely utilized to improve agriculturally important traits in Rhododendron. These interspecific hybridizations are often inhibited by reproductive barriers at the prezygotic and postzygotic stages. Thus, in order to character the barriers and reveal the underlying regulation mechanism of interspecific hybrid incompatibility in Rhododendron, we compared the pollen germination and pollen tube growth, fruit growth, seed number and germination rate between intraspecific (Rhododendron ‘XXL’ × R. ‘XXL’) and interspecific (R. ‘XXL’ × R. delavayi) hybridizations. After pollination, the pollen germination rate and pollen tube growth rate of interspecific hybridization (Inter) were always higher than that of intraspecific hybridization (Intra) during the whole observation period. Conversely, the fruits and ovules of Intra grew faster than that of Inter after 20 DAP. The ratio of shrunken embryos of Inter was 73.27 ± 10.19 %, which was significantly higher than that of Intra with 28.41 ± 8.33 % at 40 DAP. Moreover, the fruit setting rate, the average number of seeds per fruit and seed germination rate of Intra were significantly higher than that of Inter at 150 DAP. These results indicated that the barriers of interspecific incompatibility between R. ‘XXL’ × R. delavayi could be mainly ascribed to post-fertilization isolation. Consequently, RNA-Seq analysis on ovaries at three developmental stages from two hybridizations indicated that plant hormone signal transduction, flavonoid biosynthesis, MAPK signaling pathway and plant-pathogen interaction pathways might play important roles in regulation of hybrid incompatibility between R. ‘XXL’ × R. delavayi. The results can provide a basis for the study of the hybrid incompatibility mechanism of Rhododendron, and provide useful information for the breeding and conservation of genetic resources of Rhododendron.

Root growth dynamics of two southern highbush blueberry (V. corymbosum L. interspecific hybrids) cultivars in the Western Cape, South Africa

The objective of this research was to obtain a better understanding of the root growth dynamics of two southern highbush blueberry (SHB) cultivars, '8-–42′ 42′ and '9–02′, in the Western Cape, South Africa, southern hemisphere (SH). The number and length of new roots were measured from June 2020 to June 2021 in commercial orchards in the Hex River production area. Root production was similar between the cultivars and initially peaked at the onset of early winter, to coincide with flowering. A second peak occurred in mid-summer, approximately one month after fruit harvest and summer pruning (late November, SH) and continued until shoot growth declined in late autumn (early May, SH). Soil temperatures ranged between 11 and 20 °C during the first peak in root production and 17 and 31°C during the second peak, suggesting that new root production was controlled by the stage of plant development rather than soil temperature. This paper provides the first report on root development in SHB under the SH conditions and contributes towards implementing precision irrigation and fertigation strategies for evergreen systems.

Resistance of complex interspecific cotton hybrids to wilt

<p>In the article, the results of determining the degree of tolerance to wilt disease in the wilt background of hybrid families obtained using the method of interspecies complex hybridization in the cotton plant are analyzed. In the F2–F4 generations of interspecies complex hybrids, a large-scale process of separation according to the sign of tolerance to wilt disease takes place, and among the hybrids, plants that are not damaged by wilt disease, partially damaged plants, and severely damaged plants are separated. With this in mind, it was confirmed that it is possible to isolate lines and varieties with different levels of wilt damage from complex hybrid combinations of interspecies with different genomes by purposefully conducting selection work by the breeder. A number of wilt disease-tolerant families compared to control cultivars have been isolated and are recommended for use as a starting source in genetic selection studies.<strong><em></em></strong></p>

Effect of Asystasia spp. Management on the O×G Hybrid (Elaeis oleifera × Elaeis guineensis) of Oil Palm at the Adult Stage

The proper management of weeds plays an important role in nutrition and health and, therefore, in the yield and sustainability of the oil palm crop. To evaluate the effect of Asystasia spp. management as a cover species in the interspecific hybrid O×G oil palm at the adult stage, a trial was established in the Luker Agrícola plantation (Casanare, Colombia). The treatments evaluated corresponded to four types of management of Asystasia spp. with different control radii concerning the stipe (drip line) (1, 3, and 4.5 m on three occasions during the year) on an O×G Coari × La Mé cultivar at the adult stage (2011 planting). The treatments were established under a randomized complete block design with four replications. The analysis of the results obtained after four years of evaluation indicated that there are no significant differences between the treatments evaluated in terms of soil chemistry, nutrition, growth, and crop yield by analysis of variance. Nonetheless, significant differences were observed in soil cover variables and biomass and nutrient accumulation according to Tukey’s test at 5%. According to the results obtained, the management of Asystasia spp. should be focused on the conservation and efficient use of resources. It should be adapted according to the most appropriate management conditions in each case (age of the crop, weeds desired by the plantation, pests, nutrient recycling, soil moisture conservation, among others).

Integrated morphometric and molecular approaches to screen hybrid from wild Labeo rohita and Labeo catla parent populations.

Interspecific hybrids of rohu (Labeo rohita) and catla (Labeo catla) are common, especially in India due to constrained breeding. These hybrids must segregate from their wild parents as part of conservational strategies. This study intended to screen the hybrids from wild rohu and catla parents using both morphometric and molecular approaches. The carp samples were collected from Jharkhand and West Bengal, India. The correlation and regression analysis of morphometric features are considered superficial but could be protracted statistically by clustering analysis and further consolidated by nucleotide variations of one mitochondrial and one nuclear gene to differentiate hybrids from their parents. Out of 21 morphometric features, 6 were used for clustering analysis that exhibited discrete separation among rohu, catla, and their hybrids when the data points were plotted in a low-dimensional 2-D plane using the first 2 principal components. Out of 40 selected single nucleotide polymorphism (SNP) positions of the COX1 gene, hybrid showed 100% similarity with catla. Concerning SNP similarity of the 18S rRNA nuclear gene, the hybrid showed 100% similarity with rohu but not with catla; exhibiting its probable parental inheritance. Along with morphometric analysis, the SNP comparison study together points towards strong evidence of interspecific hybridization between rohu and catla, as these hybrids share both morphological and molecular differences with either parent. However, this study will help screen the hybrids from their wild parents, as a strategy for conservational management.

Asymmetric Introgression and Cryptic Natural Hybridization between Two Species of Teucrium Section Polium (Lamiaceae) on the Balkan Peninsula.

In this work, we analyzed the morphology and genetic structure of Teucrium montanum, T. capitatum and their hybrid T. × rohlenae from three syntopic populations. A morphometric study showed that the parents and their hybrids exhibited continuous morphological variation, with the hybrid positioned exactly between the parents. Genetic analysis revealed that plants morphologically identified as T. × rohlenae are fertile hybrids that produce hybrid swarms dominated by later-generation hybrids. This suggests that introgression, rather than speciation, is the more likely outcome of hybridization between these plant species. The extent and direction of gene flow between the two species differed markedly between the three syntopic localities. At the Trilj locality, it was clearly unidirectional, with T. capitatum playing the dominant role. At the Sićevo locality, gene flow was slightly asymmetric, favoring the genetic background of T. capitatum, while at the Sliven site, it was completely asymmetric in the opposite direction. The extreme case of unidirectional gene flow was observed at the Trilj locality where plants morphologically identified as T. montanum could not be genetically distinguished from T. capitatum. This suggests that interspecific hybridization occurred long ago, leading to introgression and cryptic hybrids, blurring of species boundaries and generating evolutionary noise.

Freezing Points of Fruit from Different Kiwifruit Genotypes at Harvest and during Cold Storage

Fruit storage is optimized at the lowest safe temperature to maximize storage life whilst avoiding chilling or freezing injury. The few published studies of freezing in kiwifruit (Actinidia spp.) have been conducted with A. chinensis var. deliciosa ‘Hayward’ fruit, with freezing temperatures reported in the range of −1.5 °C to −2.5 °C. In China, a large number of kiwifruit cultivars have been commercialized recently, with at least ten being commonly planted. In this research, freshly harvested fruits from 45 kiwifruit germplasm accessions were measured for freezing point, soluble solids content (SSC) and water content (WC). The difference in freezing point of different tissue zones within a fruit and after different periods of cold storage were determined for the fruit of three representative main cultivars: the green-fleshed ‘Hayward’, the red-fleshed A. chinensis var. chinensis ‘Donghong’, and the yellow-fleshed interspecific hybrid A. eriantha × A. chinensis ‘Jinyan’. The the freezing point of kiwifruit was found to be highly correlated with the SSC. This relationship was found irrespective of whether the measurement was made at harvest, at different places in the fruit, or after ripening during storage. These findings agree with previous reports for kiwifruit and other fruits, although it appears that the relationship may differ among kiwifruit cultivars, something not previously reported and which requires further confirmation.

Evolutionary genomics of two diploid goat grass species belonging to the section Sitopsis of Aegilops, Aegilops longissima, and Aegilops sharonensis.

Aegilops longissima and Ae. sharonensis, being classified into the Sitopsis section of genus Aegilops, are distinct species both taxonomically and ecologically. Nevertheless, earlier observations indicate that the two species are not reproductively isolated to full extent and can inter-bred upon secondary contact. However, the genomic underpinnings of the morpho-ecological differentiation between the two foci species remained unexplored. Here, we resequenced 31 representative accessions of the two species and conducted in-depth comparative genomic analyses. We demonstrate recurrent and ongoing natural hybridizations between Ae. longissima and Ae. sharonensis, and depict features of genome composition of the resultant hybrids at both individual and population levels. We also delineate genomic regions and candidate genes potentially underpinning the differential morphological and edaphic adaptations of the two species. Intriguingly, a binary morphology was observed in the hybrids, suggesting existence of highly diverged genomic regions that remain uneroded by the admixtures. Together, our results provide new insights into the molding effects of interspecific hybridization on genome composition and mechanisms preventing merge of the two species.

Uniparental silencing of 5S rRNA genes in plant allopolyploids - insights from Cardamine (Brassicaceae).

While the phenomenon of uniparental silencing of 35S rDNA in interspecific hybrids and allopolyploids is well documented, there is a notable absence of information regarding whether such silencing extends to the 5S RNA component of ribosomes. To address this gap in knowledge, we analyzed the 5S and 35S rDNA expression in Cardamine (Brassicaceae) allopolyploids, namely C. × insueta (2n = 3x = 24, genome composition RRA), C. flexuosa (2n = 4x = 32, AAHH), and C. scutata (2n = 4x = 32, PPAA) which share a common diploid ancestor (AA). We employed high-throughput sequencing of transcriptomes and genomes and phylogenetic analyses of 5S rRNA variants. The genomic organization of rDNA was further scrutinized through clustering and fluorescence in situ hybridization. In the C. × insueta allotriploid, we observed uniparental dominant expression of 5S and 35S rDNA loci. In the C. flexuosa and C. scutata allotetraploids, the expression pattern differed, with the 35S rDNA being expressed from the A subgenome, whereas the 5S rDNA was expressed from the partner subgenome. Both C. flexuosa and C. scutata but not C. × insueta showed copy and locus number changes. We conclude that in stabilized allopolyploids, transcription of ribosomal RNA components occurs from different subgenomes. This phenomenon appears to result in the formation of chimeric ribosomes comprising rRNA molecules derived from distinct parental origins. We speculate that the interplay of epigenetic silencing and rDNA rearrangements introduces an additional layer of variation in multimolecule ribosomal complexes, potentially contributing to the evolutionary success of allopolyploids.

The Problems of DNA-Barcoding the Shads of Genus Alosa (Alosidae) of the Ponto-Caspian Basin

—Numerous studies show that species identification of representatives of the genus Alosa using various genetic markers is often difficult and the search for more specific biomarkers is required. For the first time we analyzed polymorphism of COI gene fragment of mitochondrial DNA of two representatives of this genus (A. tanaica and A. kessleri), supplemented with new data on A. immaculata, from the waters of the Ponto-Caspian basin in comparative aspect with other representatives of the herring (Clupeoidea) genera Alosa, Clupea, Clupeonella, Sprattus, and Sardinops. The main result was the conclusion that within the genus Alosa, it is not possible to identify species using the marker used. On the one hand, specimens collected from morphologically distinct individuals and identified as different species have the same haplotypes. On the other hand, samples belonging to different species differ from each other by an insignificant number of nucleotide substitutions and do not form independent clades on the phylogram and haplotype network. This indicates the absence of genetic differentiation between the studied samples of herrings of genus Alosa into separate species and species groups when using DNA barcoding based on the COI gene. The reasons for such a phenomenon may be the following: (1) incorrect identification of species in catches, since shads (Alosidae) have high morphological flexibility and in many species, the main external morphological characters often overlap; (2) recent time of speciation by the standards of biological evolution for shads of genus Alosa; (3) difference in proportion of interspecific hybrids, which can vary significantly between populations of the same species.

UC Davis Controller Rootstocks for peach: their background and how they work

Unlike the situation with apples, until recently, there have been few commercially acceptable size-controlling rootstocks available for peach production. Nearly forty years ago a program was initiated to screen Prunus germplasm for ease of vegetative propagation, compatibility with peach and sizecontrolling potential. This initial research involved ~120 genotypes that were either various named Prunus species and cultivars or inter-specific Prunus hybrids. This collection was later narrowed down to nine selections that showed some promise for commercial production. After subsequent field trials two new rootstocks were released in 2004 (K146-43 and P30-135; Controller 5 and 9TM, respectively). Subsequently, a new breeding program was initiated based on the hypothesis that hybrids of two peach cultivars (Okinawa and Harrow Blood) would have the potential to reduce the scion vigor of peach and be resistant to root-knot nematodes. A seedling population of hybrids resulting for this cross were propagated and ‘O’Henry’ peach was grafted onto each individual genotype. Those that showed size-controlling potential were then put into a series of replicated field trials and evaluated for both size-controlling behavior and productivity characteristics. These trials resulted in the release of a series of new rootstocks (‘HBOK 27’, ‘HBOK 32’, ‘HBOK 10’, ‘HBOK 50’; Controller 6, 7, 8 and 9.5TM, respectively). Subsequent intensive research documented that a major mechanism involved in growth control of scions grafted on these rootstocks involves reduced hydraulic conductivity of these rootstocks during early spring growth caused by smaller sized xylem vessel diameters.

Genetic diversity in Pima (Gossypium barbadense L.) and advanced interspecific hybrids (Gossypium hirsutum x Gossypium barbadense) of cotton germplasm in Ethiopia

This study was conducted to assess the genetic diversity in Pima (Gossypium barbadense L.) and advanced interspecific hybrids (G. hirsutum L. x G. barbadense L.) of cotton germplasm in Ethiopia. A total of 26 germplasm were genotyped using 26 polymorphic simple sequence repeat (SSR) markers. A total of 165 polymorphic loci, with a range of 3 to 13 alleles and a mean of 6.35 per marker were detected. About 11% of total alleles were unique alleles in 11 germplasm. The gene diversity varied from 0.39 to 0. 89, while the heterozygosity was in the range of 0 to 1.00. Furthermore, the polymorphic information content of the markers varied from 0.37 (BNL1417) to 0.88 (BNL1672) with an average of 0.68. Among the complete panel of cotton germplasm used in this study, the pairwise genetic distance ranged from 0.08 to 0.94, with an overall mean of 0.61. The UPGMA cluster analysis grouped the Pima cotton germplasm into two cluster groups and the advanced interspecific hybrid cotton germplasm makes one cluster. Principal coordinate analysis indicates that the first three most informative principal coordinates explained 68.4% of the genetic variation. The result evidenced very low to high genetic dissimilarity and overall, a considerable genetic diversity among and within germplasm which could be used to further broaden the genetic base and to enlarge the number of available cotton germplasm.

Inter-specific hybrids in Areca spp.: Verification using SCoT markers

Disease resistance has been a major goal in crop improvement programmes in arecanut (Areca catechu L.), where the fruit rot diseaseor Mahali, caused by Phytophthora meadii, is a major production constraint in India. The wild Areca spp., such as A. triandra and A.concinna, have been reported to possess resistance to P. meadii. Developing inter-specific hybrids between A. catechu and A. triandraor A. concinna could be one of the strategies to introgress the disease-resistant trait from the wild Areca spp. into cultivated arecanut.In this study, we report the utilization of start codon targeted (SCoT) markers to differentiate A. catechu from A. triandra and A. concinnaand the development of sequence-characterized amplified region (SCAR) markers to enable authentication of true inter-specific hybridsbetween them. The technique would effectively verify inter-specific hybrids at the seedling stage itself.