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| Convertion and Redevelopment of Molecular Markers of 4 Pleiotropic Disease Resistance Genes in Wheat (Triticum aestivum) |
| GAO Jie, SONG Guo-Qi, LI Ji-Hu, LI Yu-Lian, ZHANG Shu-Juan, ZHANG Rong-Zhi, GU Tian-Tian, LI Gen-Ying, LI Wei* |
1 Crop Research Institute, Shandong Academy of Agricultural Sciences, Jinan 250100, China;
2 Key Laboratory of Wheat Biology and Genetic Improvement in the North Yellow & Huai River Valley, Ministry of Agriculture/National Engineering Laboratory for Wheat & Maize, Jinan 250100, China |
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Abstract It has been more than 20 years since wheat (Triticum aestivum) molecular breeding was introduced to China, only a few commercial cultivars were released. To promote combination of marker assistant selection and traditional breeding, and accelerate molecular breeding development is urgent. There are 4 well characterized pleiotropic disease resistance genes in wheat. It is difficult to select these genes in breeding program through phenotype, marker assistant selection is an option. Markers developed for pleiotropic disease resistance genes have some shortcoming, such as ambiguous genotyping or single marker type, which limit its use. In this study, Focus on the ambiguous genotyping problem of reported kompetitive allele-specific PCR (KASP) markers of pleiotropic disease resistance genes, the basic local alignment search tool (BLAST) was used to align the KASP marker amplification regions and the 'Chinese Spring' reference genome. Except the target regions, several other similar genomic regions were found. Too many similar sequences in the genome may be the reason for genotyping problem of KASP markers. According to the BLAST result, primers were redesigned. The KASP marker genotyping result of improved Lr34K2 and Lr46K3 were better than Lr34_TCCIND for leaf rust resistance gene 34 (Lr34) and Lr46_JF2-2 for Lr46, respectively; KASP markers Sr2K3 for stem rust resistance 2 (Sr2), C6K1C2 and C6K2C1 for Lr34, and Lr46g22K3 for Lr46 were redeveloped, the genotyping results were clear; KASP markers Lr46_JF2-2 for Lr46, TM4 and TM10 for Lr67 were also converted into derived cleaved amplified polymorphic sequences (dCAPS) markers Lr46Rdcaps, TM4dcaps, and TM10dcaps, respectively. dCAPS markers would release the machinery requirement and lower the application threshold. In conclusion, this study enriches marker type and quantity, and provides convenience for marker assistant selection of pleiotropic disease resistance genes. It also has important reference value for KASP marker development and improvement.
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Received: 10 August 2020
Published: 01 May 2021
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Corresponding Authors:
*davidlee5@163.com
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[1] 贾继增. 1997. 小麦分子标记研究进展[J]. 生物技术通报, 2: 1-5.
(Jia J Z1997. Review on wheat molecular markers[J]. Biotechnology Bulletin, 2: 1-5.)
[2] 李豪圣, 刘建军,宋健民,等. 2019. 优质强筋小麦济麦229选育实践与思考[J]. 山东农业科学, 51(8): 21-24.
(Li H S, Liu J J, Song J M, et al.2019. Breeding practice and consideration of wheat variety Jimai 229 with high quality and strong gluten[J]. Shandong Agricultural Sciences, 51(8): 21-24.
[3] 李玮, 宋国琦, 李吉虎, 等. 2020. 小麦四个多效抗病基因的分子检测[J]. 麦类作物学报, 40(4): 395-400.
(Li W, Song G Q, Li J H, et al.2020. Molecular detection of four pleiotropic disease resistance genes in wheat[J]. Journal of Triticeae Crops, 40(4): 395-400.)
[4] 王竹林, 刘曙东, 王辉, 等. 2006. 小麦慢病性的遗传育种研究进展[J]. 麦类作物学报, 26(1): 137-142.
(Wang Z L, Liu S D, Wang H, et al.2006. Advances of study on adult-plant resistance in bread wheat[J]. Journal of Triticeae Crops, 26(1): 137-142.)
[5] 张亚琦. 2015. 460个小麦品种抗叶锈性鉴定及Libellula成株抗叶锈QTL作图[D]. 博士学位论文, 河北农业大学, 导师: 李在峰, 刘大群, pp.12.
(Zhang Y Q.2015. Identifcation of wheat leaf rust resistance genes in 460 Chinese wheat cultivars and QTL mapping for adult-plant resistance gene to leaf rust in Italian wheat cultivar Libellula[D]. Thesis for Ph.D., Agricultural University of Hebei, Supervisor: Li Z F, Liu D Q, pp. 12.)
[6] Dakouri A, McCallum B D, Walichnowski A Z, et al.2010. Fine-mapping of the leaf rust Lr34 locus in Triticum aestivum (L.) and characterization of large germplasm collections support the ABC transporter as essential for gene function[J]. Theoretical and Applied Genetics, 121(2): 373-384.
[7] Fang T, Lei L, Li G, et al.2020. Development and deployment of KASP markers for multiple alleles of Lr34 in wheat[J]. Theoretical and Applied Genetics: 1-13.
[8] Herrera-Foessel S A, Singh R P, Lillemo M, et al.2014. Lr67/Yr46 confers adult plant resistance to stem rust and powdery mildew in wheat[J]. Theoretical and Applied Genetics, 127(4): 781-789.
[9] Krattinger S G, Jordan D R, Mace E S, et al.2013. Recent emergence of the wheat Lr34 multi-pathogen resistance: insights from haplotype analysis in wheat, rice, sorghum and Aegilops tauschii[J]. Theoretical and Applied Genetics, 126(3): 663-672.
[10] Lagudah E S, Krattinger S G, Herrerafoessel S, et al.2009. Gene-specific markers for the wheat gene Lr34/Yr18/Pm38 which confers resistance to multiple fungal pathogens[J]. Theoretical and Applied Genetics, 119(5): 889-898.
[11] Lagudah E S, Mcfadden H, Singh R P, et al.2006. Molecular genetic characterization of the Lr34/Yr18 slow rusting resistance gene region in wheat[J]. Theoretical andApplied Genetics, 114(1): 21-30.
[12] Lan C, Rosewarne G M, Singh R P, et al.2014. QTL characterization of resistance to leaf rust and stripe rust in the spring wheat line Francolin#1[J]. Molecular Breeding, 34(3): 789-803.
[13] Liu Y, He Z, Appels R, et al.2012. Functional markers in wheat: Current status and future prospects[J]. Theoretical and Applied Genetics, 125(1): 1-10.
[14] Mago R, Simkova H, Brown-Guedira G, et al.2011. An accurate DNA marker assay for stem rust resistance gene Sr2 in wheat[J]. Theoretical and Applied Genetics, 122(4): 735-744.
[15] Mago R, Tabe L, Vautrin S, et al.2014. Major haplotype divergence including multiple germin-like protein genes, at the wheat Sr2 adult plant stem rust resistance locus[J]. BMC Plant Biology, 14: 379.
[16] Makhoul M, Rambla C, Voss-Fels K P, et al.2020. Overcoming polyploidy pitfalls: A user guide for effective SNP conversion into KASP markers in wheat[J]. Theoretical and Applied Genetics, 133(8): 2413-2430.
[17] Moore J W, Herrera-Foessel S, Lan C, et al.2015. A recently evolved hexose transporter variant confers resistance to multiple pathogens in wheat[J]. Nature Genetics, 47(12): 1494-1498.
[18] Neff M M, Turk E, Kalishman M.2002. Web-based primer design for single nucleotide polymorphism analysis[J]. Trends in Genetics, 18(12): 613-615.
[19] Rasheed A, Wen W, Gao F, et al.2016. Development and validation of KASP assays for genes underpinning key economic traits in bread wheat[J]. Theoretical and Applied Genetics, 129(10): 1843-1860.
[20] Tabe L, Samuel S, Dunn M, et al.2019. The phenotypes conferred by wheat multiple pathogen resistance locus, Sr2, include cell death in response to biotic and abiotic stresses[J]. Phytopathology, 109(10): 1751-1759. |
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