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Keywords = marker-assisted breeding (MAB)

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24 pages, 1667 KB  
Article
ddRADseq Applications for Petunia × hybrida Clonal Line Breeding: Genotyping and Variant Identification for Target-Specific Assays
by Angelo Betto, Francesco Scariolo, Giovanni Gabelli, Damiano Riommi, Silvia Farinati, Alessandro Vannozzi, Fabio Palumbo and Gianni Barcaccia
Horticulturae 2026, 12(2), 160; https://doi.org/10.3390/horticulturae12020160 - 30 Jan 2026
Viewed by 727
Abstract
Molecular genotyping is a key factor for plant breeding programming and plant variety protection (PVP). However, its potential still remains to be elucidated when considering ornamental plants like Petunia × hybrida. In this study, a petunia breeding clone collection, including sister line [...] Read more.
Molecular genotyping is a key factor for plant breeding programming and plant variety protection (PVP). However, its potential still remains to be elucidated when considering ornamental plants like Petunia × hybrida. In this study, a petunia breeding clone collection, including sister line groups, was genotyped through double digest Restriction-site Associated DNA sequencing (ddRADseq), and its genetic diversity and structure were studied. In addition to estimating the high genetic similarity observed among sister lines, this approach allowed the unique discrimination of each clone too. Molecular results agreed with genealogy data, supporting the assessment of genotyping effectiveness. In addition, the minimal number of variants able to uniquely discriminate and/or correctly cluster the experimental lines was investigated. The loci number could be reduced to eight to achieve line discrimination, and a method to identify the specific variant sets is presented. Conversely, to preserve the original clustering with minor adjustments, one hundred loci were required and were obtained through minor allele frequency (MAF) filtering. Moreover, analysis of the chromosomal distribution of variants revealed a predominant accumulation in distal regions. Genetic analyses were repeated considering only variants located in coding sequences and results were in agreement with what previously observed, disclosing the potential of the expressed regions for genotyping purposes. Eventually, the applied approach enabled the investigation of SNPs within genes putatively involved in traits of interest. Our findings encourage the adoption of high-throughput and cost-effective sequencing techniques for petunia genotyping aimed at achieving PVP, supporting new variety registration, and developing marker-assisted breeding (MAB) and marker-assisted selection (MAS) strategies. Full article
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16 pages, 3749 KB  
Article
Genotyping of Commercial European Cannabis Seeds Based on Multiple Mapped Marker Loci: A Comparative Study of Drug and Hemp Varieties
by Marcello Borin, Francesco Scariolo, Maddalena Cappello Fusaro, Irene Lucchetta, Gio Batta Sacilotto, Marco Gazzola, Stefano Bona and Gianni Barcaccia
Plants 2025, 14(19), 3050; https://doi.org/10.3390/plants14193050 - 2 Oct 2025
Cited by 1 | Viewed by 1545
Abstract
Cannabis sativa L. (2n = 2x = 20) is a widely recognized species within the Cannabaceae family. Despite its utilization for medicinal, recreational, and industrial purposes, alongside its extensive historical background, the number of genetic and biotechnological studies of this plant species has [...] Read more.
Cannabis sativa L. (2n = 2x = 20) is a widely recognized species within the Cannabaceae family. Despite its utilization for medicinal, recreational, and industrial purposes, alongside its extensive historical background, the number of genetic and biotechnological studies of this plant species has decreased due to legal ramifications and prohibition campaigns associated with its use and cultivation. For many years, the development of novel varieties has been pursued solely by cultivators, as domestic growers have transitioned their work from cultivation to breeding Cannabis lineages. Recently, the application of genomics has facilitated a surge in methodologies aimed at marker-assisted selection, germplasm management, genetic differentiation, authentication of cultivated varieties or cultivars, and forensic applications such as safeguarding intellectual property rights. Nevertheless, the utilization of molecular markers for the advancement of commercial varieties through marker-assisted breeding (MAB) frameworks remains rare. This investigation was designed to evaluate a previously established informative microsatellite (SSR) array for the genotyping of drug-type Cannabis sativa cultivars derived from seeds of European origin. A total of 171 samples from 20 varieties were collected from European distributors and analyzed for genetic uniformity and population structure. The results were then compared with previously analyzed hemp samples and drug-type samples of Canadian origin, revealing the identification capabilities of our SSR genotyping method. Full article
(This article belongs to the Section Plant Genetics, Genomics and Biotechnology)
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22 pages, 2609 KB  
Review
Genome-Wide Datasets of Chicories (Cichorium intybus L.) for Marker-Assisted Crop Breeding Applications: A Systematic Review and Meta-Analysis
by Samela Draga, Giovanni Gabelli, Fabio Palumbo and Gianni Barcaccia
Int. J. Mol. Sci. 2023, 24(14), 11663; https://doi.org/10.3390/ijms241411663 - 19 Jul 2023
Cited by 8 | Viewed by 3543
Abstract
Cichorium intybus L. is the most economically important species of its genus and among the most important of the Asteraceae family. In chicory, many linkage maps have been produced, several sets of mapped and unmapped markers have been developed, and dozens of genes [...] Read more.
Cichorium intybus L. is the most economically important species of its genus and among the most important of the Asteraceae family. In chicory, many linkage maps have been produced, several sets of mapped and unmapped markers have been developed, and dozens of genes linked to traits of agronomic interest have been investigated. This treasure trove of information, properly cataloged and organized, is of pivotal importance for the development of superior commercial products with valuable agronomic potential in terms of yield and quality, including reduced bitter taste and increased inulin production, as well as resistance or tolerance to pathogens and resilience to environmental stresses. For this reason, a systematic review was conducted based on the scientific literature published in chicory during 1980–2023. Based on the results obtained from the meta-analysis, we created two consensus maps capable of supporting marker-assisted breeding (MAB) and marker-assisted selection (MAS) programs. By taking advantage of the recently released genome of C. intybus, we built a 639 molecular marker-based consensus map collecting all the available mapped and unmapped SNP and SSR loci available for this species. In the following section, after summarizing and discussing all the genes investigated in chicory and related to traits of interest such as reproductive barriers, sesquiterpene lactone biosynthesis, inulin metabolism and stress response, we produced a second map encompassing 64 loci that could be useful for MAS purposes. With the advent of omics technologies, molecular data chaos (namely, the situation where the amount of molecular data is so complex and unmanageable that their use becomes challenging) is becoming far from a negligible issue. In this review, we have therefore tried to contribute by standardizing and organizing the molecular data produced thus far in chicory to facilitate the work of breeders. Full article
(This article belongs to the Special Issue Genetics and Multi-Omics for Crop Breeding)
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26 pages, 9712 KB  
Article
Pyramiding Submergence Tolerance and Three Bacterial Blight Resistance Genes in Popular Rice Variety Hasanta through Marker-Assisted Backcross Breeding
by Goutam Das, Banshidhar Pradhan, Debendranath Bastia, Sanghamitra Samantaray, Debarchana Jena, Diptibala Rout, Paduranga Bhagwan Arsode, Vineeta Singh, Arup Kumar Mukherjee, Chander Mohan and Ramlakhan Verma
Agriculture 2022, 12(11), 1815; https://doi.org/10.3390/agriculture12111815 - 31 Oct 2022
Cited by 10 | Viewed by 4669
Abstract
Flash flooding (FF) along with bacterial blight (BB) outbreak are very destructive for rice production in the rainfed shallow-lowland (RSL) ecosystem. The presence of dynamic Xoo races with varying levels of genetic diversity and virulence renders their management extremely challenging under RSL. In [...] Read more.
Flash flooding (FF) along with bacterial blight (BB) outbreak are very destructive for rice production in the rainfed shallow-lowland (RSL) ecosystem. The presence of dynamic Xoo races with varying levels of genetic diversity and virulence renders their management extremely challenging under RSL. In this context, the marker-assisted improvement of plant resistance/tolerance has been proven as one of the most promising strategies towards the development of sustainable cultivars. The present study demonstrates the marker-assisted introgression of the submergence tolerant (Sub1) and three bacterial blight resistant genes (Xa21 + xa13 + xa5) into the genetic background of Hasanta, a long duration popular rice variety in the eastern coastal region of India. The rice genotypes, Swarna Sub1 (carrying Sub1) and IRBB66 (carrying Xa21 + xa13 + Xa7 + xa5 + Xa4) had maximum genetic similarity (0.84 and 0.73, respectively) with Hasanta; recurrent parent (RP) was used as donor. The forward analysis of target genes in F1s, IC1F1s and backcross (BC) generations was performed by linked/genic markers (Sub1bc2; pTA248, xa13prom and RM122), whereas background recovery of RP in each BC and segregating generations was performed utilizing 108 hypervariable SSR markers. Intervened speed breeding (SB) strategy and intensive phenotyping could lead the development of near isogenic lines (NILs) as to the RP in all basic traits. The performance of the near isogenic lines (NILs, BC2F3 and BC2F4), HS 232-411-391-756-37, HS 232-411-391-809-8, HS 232-411-391-756-18, HS 110-224-197-10-36, HS 232-411-391-809-81, HS 110-224-197-10-41 and HS 232-411-391-809-63 establishes the utility of marker-assisted backcross-breeding (MAB) and SB in accelerated trait introgression. The introgressed lines carrying Sub1 + Xa21 + xa13 + xa5 showed 76% to 91% survival under 14 days of submergence and durable BB resistance (percent disease index-PDI of 2.68 ± 0.26 to 6.22 ± 1.08 and lesion length, LL of 1.29 ± 0.12 to 4.2 ± 0.64 cm). Physiological analysis revealed that improved NILs, carrying Sub1 gene conquered adaptive physiological modulations, had reduced the consumption of soluble sugar and the degradation of total chlorophyll contents (TCC), and an enhanced level of Alcohol Dehydrogenase activity (ADH) and proline accumulation in all submergence regimes. The pyramided lines attained complete product profile of RP, that will contribute to sustainable rice production under RSL, particularly in the coastal region that has substantial acreage under the variety Hasanta. Full article
(This article belongs to the Special Issue Prospects and Challenges of Rice Breeding under Climate Change)
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14 pages, 1842 KB  
Article
Submergence Gene Sub1A Transfer into Drought-Tolerant japonica Rice DT3 Using Marker-Assisted Selection
by Yong-Pei Wu, Shu-Mei Wang, Yu-Chi Chang, Chi Ho and Yu-Chia Hsu
Int. J. Mol. Sci. 2021, 22(24), 13365; https://doi.org/10.3390/ijms222413365 - 13 Dec 2021
Cited by 11 | Viewed by 4286
Abstract
Flash flooding is a major environmental stressor affecting rice production worldwide. DT3 is a drought-tolerant, recurrent parent with a good yield, edible quality, and agronomic traits akin to those of an elite Taiwanese variety, Taiken9 (TK9). Progenies carrying Sub1A can enhance submergence stress [...] Read more.
Flash flooding is a major environmental stressor affecting rice production worldwide. DT3 is a drought-tolerant, recurrent parent with a good yield, edible quality, and agronomic traits akin to those of an elite Taiwanese variety, Taiken9 (TK9). Progenies carrying Sub1A can enhance submergence stress tolerance and can be selected using the marker-assisted backcross (MAB) breeding method. For foreground selection, Sub1A and SubAB1 were utilized as markers on the BC2F1, BC3F1, and BC3F2 generations to select the submergence-tolerant gene, Sub1A. Background selection was performed in the Sub1A-BC3F2 genotypes, and the percentages of recurrent parent recovery within individuals ranged from 84.7–99.55%. BC3F3 genotypes (N = 100) were evaluated for agronomic traits, yield, and eating quality. Four of the eleven BC3F4 lines showed good yield, yield component, grain, and eating quality. Four BC3F4 lines, SU39, SU40, SU89, and SU92, exhibited desirable agronomic traits, including grain quality and palatability, consistent with those of DT3. These genotypes displayed a high survival rate between 92 and 96%, much better compared with DT3 with 64%, and demonstrated better drought tolerance compared to IR64 and IR96321-345-240. This study provides an efficient and precise MAB strategy for developing climate-resilient rice varieties with good grain quality for flood-prone regions. Full article
(This article belongs to the Special Issue Molecular Research in Rice: Agronomically Important Traits 2.0)
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41 pages, 4438 KB  
Review
Genomic Approaches for Improvement of Tropical Fruits: Fruit Quality, Shelf Life and Nutrient Content
by Malarvizhi Mathiazhagan, Bhavya Chidambara, Laxman R. Hunashikatti and Kundapura V. Ravishankar
Genes 2021, 12(12), 1881; https://doi.org/10.3390/genes12121881 - 25 Nov 2021
Cited by 57 | Viewed by 10097
Abstract
The breeding of tropical fruit trees for improving fruit traits is complicated, due to the long juvenile phase, generation cycle, parthenocarpy, polyploidy, polyembryony, heterozygosity and biotic and abiotic factors, as well as a lack of good genomic resources. Many molecular techniques have recently [...] Read more.
The breeding of tropical fruit trees for improving fruit traits is complicated, due to the long juvenile phase, generation cycle, parthenocarpy, polyploidy, polyembryony, heterozygosity and biotic and abiotic factors, as well as a lack of good genomic resources. Many molecular techniques have recently evolved to assist and hasten conventional breeding efforts. Molecular markers linked to fruit development and fruit quality traits such as fruit shape, size, texture, aroma, peel and pulp colour were identified in tropical fruit crops, facilitating Marker-assisted breeding (MAB). An increase in the availability of genome sequences of tropical fruits further aided in the discovery of SNP variants/Indels, QTLs and genes that can ascertain the genetic determinants of fruit characters. Through multi-omics approaches such as genomics, transcriptomics, metabolomics and proteomics, the identification and quantification of transcripts, including non-coding RNAs, involved in sugar metabolism, fruit development and ripening, shelf life, and the biotic and abiotic stress that impacts fruit quality were made possible. Utilizing genomic assisted breeding methods such as genome wide association (GWAS), genomic selection (GS) and genetic modifications using CRISPR/Cas9 and transgenics has paved the way to studying gene function and developing cultivars with desirable fruit traits by overcoming long breeding cycles. Such comprehensive multi-omics approaches related to fruit characters in tropical fruits and their applications in breeding strategies and crop improvement are reviewed, discussed and presented here. Full article
(This article belongs to the Special Issue Molecular Breeding Approaches in Tropical Horticulture Species)
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19 pages, 4364 KB  
Article
A New Breeding Strategy towards Introgression and Characterization of Stay-Green QTL for Drought Tolerance in Sorghum
by Nasrein Mohamed Kamal, Yasir Serag Alnor Gorafi, Hanan Abdeltwab, Ishtiag Abdalla, Hisashi Tsujimoto and Abdelbagi Mukhtar Ali Ghanim
Agriculture 2021, 11(7), 598; https://doi.org/10.3390/agriculture11070598 - 28 Jun 2021
Cited by 10 | Viewed by 3910
Abstract
Several marker-assisted selection (MAS) or backcrossing (MAB) approaches exist for polygenic trait improvement. However, the implementation of MAB remains a challenge in many breeding programs, especially in the public sector. In MAB introgression programs, which usually do not include phenotypic selection, undesired donor [...] Read more.
Several marker-assisted selection (MAS) or backcrossing (MAB) approaches exist for polygenic trait improvement. However, the implementation of MAB remains a challenge in many breeding programs, especially in the public sector. In MAB introgression programs, which usually do not include phenotypic selection, undesired donor traits may unexpectedly turn up regardless of how expensive and theoretically powerful a backcross scheme may be. Therefore, combining genotyping and phenotyping during selection will improve understanding of QTL interactions with the environment, especially for minor alleles that maximize the phenotypic expression of the traits. Here, we describe the introgression of stay-green QTL (Stg1–Stg4) from B35 into two sorghum backgrounds through an MAB that combines genotypic and phenotypic (C-MAB) selection during early backcross cycles. The background selection step is excluded. Since it is necessary to decrease further the cost associated with molecular marker assays, the costs of C-MAB were estimated. Lines with stay-green trait and good performance were identified at an early backcross generation, backcross two (BC2). Developed BC2F4 lines were evaluated under irrigated and drought as well as three rainfed environments varied in drought timing and severity. Under drought conditions, the mean grain yield of the most C-MAB-introgression lines was consistently higher than that of the recurrent parents. This study is one of the real applications of the successful use of C-MAB for the development of drought-tolerant sorghum lines for drought-prone areas. Full article
(This article belongs to the Special Issue Biotic and Abiotic Stresses in Crop Plants)
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8 pages, 1698 KB  
Article
Evaluating Multiple Allelic Combination to Determine Tiller Angle Variation in Rice
by Su Jang, Yoo Seok Kang, Yoon Kyung Lee and Hee-Jong Koh
Agriculture 2020, 10(10), 428; https://doi.org/10.3390/agriculture10100428 - 25 Sep 2020
Cited by 2 | Viewed by 3606
Abstract
Tiller angle is an important influencing factor in rice plant architecture that affects planting density and yield per unit area. Molecular tools to predict tiller angle contribute to breeding programs, which aim at optimizing rice plant architecture. In this study, several single-nucleotide polymorphism [...] Read more.
Tiller angle is an important influencing factor in rice plant architecture that affects planting density and yield per unit area. Molecular tools to predict tiller angle contribute to breeding programs, which aim at optimizing rice plant architecture. In this study, several single-nucleotide polymorphism (SNP) markers related to tiller angle were developed and used with a model population to define a linear regression model for the prediction of tiller angle in rice. The resulting linear regression model, consisting of eight SNP markers as independent variables, was assessed using an independent test population. Overall, the regression model achieved an adjusted R2 of 0.51 and exhibited consistent predictive accuracy with an R2 of 0.61. Three of the eight independent variables, namely, PIN2-1, LIC1-1, and TAC1, contributed substantially to the linear regression model. These three major effect markers were also major determinants of tiller angle in the independent test population. Allelic combinations of the three major effect markers modulated tiller angle in the range of 5.6–19°. The DNA markers and linear regression model developed in this study will facilitate rice breeding programs for improving plant architecture. Full article
(This article belongs to the Special Issue Rice Breeding and Genetics)
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17 pages, 1289 KB  
Article
Improvement of a RD6 Rice Variety for Blast Resistance and Salt Tolerance through Marker-Assisted Backcrossing
by Korachan Thanasilungura, Sukanya Kranto, Tidarat Monkham, Sompong Chankaew and Jirawat Sanitchon
Agronomy 2020, 10(8), 1118; https://doi.org/10.3390/agronomy10081118 - 1 Aug 2020
Cited by 22 | Viewed by 5819
Abstract
RD6 is one of the most favorable glutinous rice varieties consumed throughout the north and northeast of Thailand because of its aroma and softness. However, blast disease and salt stress cause decreases in both yield quantity and quality during cultivation. Here, gene pyramiding [...] Read more.
RD6 is one of the most favorable glutinous rice varieties consumed throughout the north and northeast of Thailand because of its aroma and softness. However, blast disease and salt stress cause decreases in both yield quantity and quality during cultivation. Here, gene pyramiding via marker-assisted backcrossing (MAB) using combined blast resistance QTLs (qBl 1, 2, 11, and 12) and Saltol QTL was employed in solving the problem. To pursue our goal, the RD6 introgression line (RGD07005-12-165-1), containing four blast-resistant QTLs, were crossed with the Pokkali salt tolerant variety. Blast resistance evaluation was thoroughly carried out in the fields, from BC2F2:3 to BC4F4, using the upland short-row and natural field infection methods. Additionally, salt tolerance was validated in both greenhouse and field conditions. We found that the RD6 “BC4F4 132-12-61” resulting from our breeding programme successfully resisted blast disease and tolerated salt stress, while it maintained the desirable agronomic traits of the original RD6 variety. This finding may provide a new improved rice variety to overcome blast disease and salt stress in Northeast Thailand. Full article
(This article belongs to the Special Issue Molecular Marker Technology for Crop Improvement)
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36 pages, 3754 KB  
Article
High-Resolution Mapping in Two RIL Populations Refines Major “QTL Hotspot” Regions for Seed Size and Shape in Soybean (Glycine max L.)
by Aiman Hina, Yongce Cao, Shiyu Song, Shuguang Li, Ripa Akter Sharmin, Mahmoud A. Elattar, Javaid Akhter Bhat and Tuanjie Zhao
Int. J. Mol. Sci. 2020, 21(3), 1040; https://doi.org/10.3390/ijms21031040 - 4 Feb 2020
Cited by 75 | Viewed by 6493
Abstract
Seed size and shape are important traits determining yield and quality in soybean. However, the genetic mechanism and genes underlying these traits remain largely unexplored. In this regard, this study used two related recombinant inbred line (RIL) populations (ZY and K3N) evaluated in [...] Read more.
Seed size and shape are important traits determining yield and quality in soybean. However, the genetic mechanism and genes underlying these traits remain largely unexplored. In this regard, this study used two related recombinant inbred line (RIL) populations (ZY and K3N) evaluated in multiple environments to identify main and epistatic-effect quantitative trait loci (QTLs) for six seed size and shape traits in soybean. A total of 88 and 48 QTLs were detected through composite interval mapping (CIM) and mixed-model-based composite interval mapping (MCIM), respectively, and 15 QTLs were common among both methods; two of them were major (R2 > 10%) and novel QTLs (viz., qSW-1-1ZN and qSLT-20-1K3N). Additionally, 51 and 27 QTLs were identified for the first time through CIM and MCIM methods, respectively. Colocalization of QTLs occurred in four major QTL hotspots/clusters, viz., “QTL Hotspot A”, “QTL Hotspot B”, “QTL Hotspot C”, and “QTL Hotspot D” located on Chr06, Chr10, Chr13, and Chr20, respectively. Based on gene annotation, gene ontology (GO) enrichment, and RNA-Seq analysis, 23 genes within four “QTL Hotspots” were predicted as possible candidates, regulating soybean seed size and shape. Network analyses demonstrated that 15 QTLs showed significant additive x environment (AE) effects, and 16 pairs of QTLs showing epistatic effects were also detected. However, except three epistatic QTLs, viz., qSL-13-3ZY, qSL-13-4ZY, and qSW-13-4ZY, all the remaining QTLs depicted no main effects. Hence, the present study is a detailed and comprehensive investigation uncovering the genetic basis of seed size and shape in soybeans. The use of a high-density map identified new genomic regions providing valuable information and could be the primary target for further fine mapping, candidate gene identification, and marker-assisted breeding (MAB). Full article
(This article belongs to the Special Issue Legume Genetics and Biology: From Mendel's Pea to Legume Genomics)
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