Screeninfg Of Fine Tomato Inbred Lines And Development Of KASP Molecular Markers For Disease Resistance Gene

Tomato is one of the most valuable horticultural crop in the world for its strong adaptability,highly yield,good economic feedback,delicious flavor,and fertile nutritional value,at the same time it’s also a crop that sensitive to various diseases.There are more than 40 diseases occour in tomato cultivation process.Accompany with the area expanding of the protected environment,continuous croping and other negative elements,the occourence of the tomato diseases has becoming more and more serious,as a result,accelerating new tomato varieties breeding in multiple diseases resistant genes aggregation is the most economical and effective solution in logical.In this study,we used molecular markers to detect diseases resistance genes and field agronomic traits to screen high performance such as resistance to major diseases and good economic characters within tomato inbred lines.We evaluated reported KASP molecular markers and developed three KASP molecular resistance gene markers.The main research results are as follows:1.Molecular marker detection and screening of disease resistance inbred lines:we purified and screened 49 F₅ generation inbred lines in spring of 2022 and obtained 125inbred lines,among which,got 67 excellent inbred lines.On he following autumn,we found158 individual inbred lines based on the 67 inbred lines originated in the spring of 2022.By comprehensively analyzing the statistical results of the two aspects mentioned above,we seleced 84 excellent individual inbred lines for further screening.In 2023,we obtained one single inbred line named 22TA144 which with good comprehensive performance and resistance genes of five diseases on homozygous level.22TA144 could be used as an excellent father material apply to the hybrid combination in next steps.2.Development and verification of KASP molecular markers:firstly,we varified two KASP molecular markers based on Ty-1 and Sw-5 genes can accurately genotyping our Laboratory’s tomato breeding material according to previous studies;secondly,we designed three applicable KASP molecular markers independently.These five KASP molecular markers can distinguish materials’s disease-resistant or susceptible effectively,accelerated the breeding process.Based to the genomic information of Tomato Verticillium Wilt（TVW） resistance gene Ve1,tomato Tobacco Mosaic Virus（TMV）disease resistance gene Tm-2and Fusarium Crown Root and Rot（FCRR）resistance gene Frl,we designed KASP primers for specific SNP sites of these genes.We selected samples from breeding materials of different genetic backgrounds for KASP genotyping verification and gel electrophoresis molecular marker detection randomly in our lab.By comparing analysis,we found that these five molecular all have 100%consistent with the results of gel electrophoresis.Above results ensured the applicability of all five KASP markers in the subsequent detection of diseases resistance genes on tomato inbred lines.