Validation of some disease-resistance molecular markers associated with multiple diseases in tomato for marker-assisted selection program

Nevame, Adedze Yawo Mawunyo; Xia, Lu; Zhang, Wenting; Nchongboh, Chofong Gilbert; Wen-hu, LI; Hasan, Muhammad Mahmudul; Alam, Md. Amirul; Long-ting, SI

doi:10.2306/scienceasia1513-1874.2020.006

Cited by 3 publications

(2 citation statements)

References 15 publications

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“…Для апробации маркера было проведено его тестирование на 345 гибридах томата F 2 , полученных от скрещивания двух контрастных форм [26]. Также данный маркер был успешно использован при анализе 964 линий томатов разных семеноводческих компаний Китая [27].…”

Section: результаты и обсужденияunclassified

Creation of new tomato forms with fungal disease resistance genes based on marker selection

et al. 2021

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Relevance. The presented studies are aimed at obtaining new forms of tomato with a complex of genes for resistance to fungal diseases in combination with a standard type of bush and dark coloring of fruits based on marker-mediated selection.Methodology. The biological objects of the study are varieties and hybrid forms of tomato from the collection of the Michurinsky SAU. Molecular genetic analysis was performed using the following methods. DNA extraction was carried out from young leaves using a kit for isolation of NC Sample NC manufactured by Agrodiagnostika LLC according to the manufacturer's protocol. Fermentas production kits were used for PCR. Identification of the cladosporosis resistance gene was Cf-19 performed using the DNA marker R7. The presence of a fusarious wilting resistance gene was determined by a I-2/5 marker. The amplification results were visualized by agarose gel electrophoresis.Results. During the research, a collection of varieties and hybrid forms of tomato of the Michurinsky GAU was analyzed in order to identify genes for resistance to cladosporiosis Cf-19 and fusarium wilt I-2. A total of 52 genotypes were analyzed. It was found that most samples (41 samples) are characterized by a heterozygous state of the Cf-19 gene. All indeterminant and semi-determinant forms had both alleles. Of the 23 determinant forms presented in the collection, 10 had only one allele corresponding to recessive homozygote. Among all analyzed tomato genotypes, no dominant homozygous forms were noted. The study of the collection revealed several alleles of the I-2 gene. In total, four fragments corresponding to various alleles were amplified. A total of 50 resistant genotypes have been identified in the collection. Two alleys of the I-2 gene (633/693 bp) were identified in 42 tomato samples. Four varieties are homozygous in one allele (633 bp), which determines resistance. Three varieties have a second resistance allele (566 bp). One genotype has only an allele defining susceptibility (693 bp). On the basis of molecular analysis, as well as an assessment of the type of bush and fetal color, initial forms were selected with subsequent hybridization. 67 hybrid tomato plants were obtained. Evaluation of the presence of resistance genes showed that most of the resulting hybrids are resistant to cladosporiosis and fuzariosis. This is due to the presence of dominant alleles of Cf-19 and I-2 genes in a heterozygous state. Among the resulting hybrids, plants with a bark type of bush were identified. A total of 13 such plants were obtained.Conclusion. Thus, the work carried out allowed to obtain hybrid forms of tomato combine the signs of resistance to two pathogens of fungal diseases and the stem type of the bush. These forms are planned to be used in further selection work.

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Section: результаты и обсужденияunclassified

Creation of new tomato forms with fungal disease resistance genes based on marker selection

et al. 2021

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“…Current breeding studies are aimed to develop F 1 hybrids that have a higher fruit yield, quality and disease resistance (Liu et al 2014;Miyatake et al 2016;Rakha et al 2020Rakha et al , 2021. During the breeding process, genotyping and selection based on DNA markers provide substantial advantages over traditional phenotypic screening alone (Nevamea et al 2020). Studies on the identification of genes that provide tolerance to biotic and abiotic stresses and their usage in eggplant breeding programmes have been discussed for many years (Chapman 2020;Toppino et al 2021).…”

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Screening and validation of three molecular markers for disease resistance in eggplant

Sülü¹,

Polat²,

Boyaci³

et al. 2022

CZECH J GENET PLANT

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Fusarium oxysporum Schlecht. f.sp. melongenae (FOM), Verticillium dahliae (Ve) and Ralstonia solanacearum are major limiting pathogens affecting eggplant cultivation and their yield in the world. Natural resistance genes are the most environmentally friendly method to control the disease. Thus, marker-assisted selection (MAS) is preferred as a tool for screening resistance genes (R-genes) in eggplant resistance breeding. In this study, markers that are specifically linked to major disease resistance genes conferring resistance to F. melongenae, V. dahliae and R. solanacearum were tested in a population containing breeding materials to validate the resistance. Resistant Solanum melongena accessions LS1934 and LS2436 and their reciprocal crosses were used as the resistance resource for this validation study. Moreover, classical resistance tests to FOM and Ve were performed with the root-dip inoculation method for classification of all the accessions based on their resistance/susceptibility responses. The SCAR<sub>426</sub>, CAPs_903 and SIVR844 markers were highly informative for the determination of resistance genes (Fomg, Ve and ERs1). Therefore, in areas with high susceptibility to diseases, a highly efficient combination of the relevant R-genes and their pyramiding into commercial eggplant varieties are proposed to be implemented as a pragmatic approach.

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Response Surface Methodology for Optimization of Multiplex-PCR Protocols for Detection of TYLCV, TSWV and Fol Molecular Markers: Analytical Performance Evaluation

Lafrance

Valdez-Torres

Villicaña

et al. 2023

Genes

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Molecular markers linked to disease resistance genes which affect economically important crops are of great interest. In the case of tomato, a major focus on resistance breeding to multiple fungal and viral pathogens such as Tomato yellow leaf curl virus (TYLCV), Tomato spotted wilt virus (TSWV) and Fusarium oxysporum f. sp. lycopersici (Fol), have led to the introgression of several resistance genes; therefore, molecular markers have become important in molecular-assisted selection (MAS) of tomato varieties resistant to those pathogens. However, assays that allow simultaneous evaluation of resistant genotypes, such as multiplex PCR, need to be optimized and evaluated to demonstrate their analytical performance, as many factors can affect them. This work aimed to generate multiplex PCR protocols for the joint detection of the molecular markers associated with pathogen resistance genes in tomato plants that are sensitive, specific and repeatable. For the optimization a central composite design of a response surface methodology (RSM-CCD) was used. For analytical performance evaluation, specificity/selectivity and sensibility (limit of detection and dynamic range) were analyzed. Two protocols were optimized: the first one with a desirability of 1.00, contained two markers (At-2 and P7-43) linked to I- and I-3-resistant genes. The second one with a desirability of 0.99, contained markers (SSR-67, SW5 and P6-25) linked to I-, Sw-5-, and Ty-3-resistant genes. For protocol 1, all the commercial hybrids (7/7) were resistant to Fol, and for protocol 2, two hybrids were resistant to Fol, one to TSWV and one to TYLCV with good analytical performance. In both protocols, the varieties considered susceptible to the pathogens, no-amplicon or susceptible amplicons, were observed. The optimized multiplex PCR protocols showed dynamic ranges from 5.97 up to 161.3 ng DNA. The limit of detection was 17.92 ng and 53.76 ng DNA for protocols 1 and 2, respectively, giving 100% positive results in the test replicates. This method allowed to develop optimized multiplex PCR protocols with few assays which translates into less time and resources, without sacrificing method performance.

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Validation of some disease-resistance molecular markers associated with multiple diseases in tomato for marker-assisted selection program

Cited by 3 publications

References 15 publications

Creation of new tomato forms with fungal disease resistance genes based on marker selection

Creation of new tomato forms with fungal disease resistance genes based on marker selection

Screening and validation of three molecular markers for disease resistance in eggplant

Response Surface Methodology for Optimization of Multiplex-PCR Protocols for Detection of TYLCV, TSWV and Fol Molecular Markers: Analytical Performance Evaluation

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