|
|
|
| Precise Genotyping of eIF(iso)4E.a and eIF(iso)4E.c in Main Chinese Cabbage (Brassica rapa ssp. pekinensis) Cultivars and Germplasms |
|
|
|
|
Abstract Abstract Virus disease caused by Turnip mosaic virus (TuMV) is one of the most serious problems during Chinese cabbage (Brassica rapa ssp. pekinensis) cultivation. Initial infection of TuMV in Chinese cabbage has been demonstrated to be related to wild type BraA.eIF(iso)4E.a (A) and BraA.eIF(iso)4E.c (C). Loss-of-function mutation in one or two genes hinders TuMV infection and results in virus-resistant phynotype in Chinese cabbage. Two kind of allelic variations, miss splicing mutation (a1) and psedogene mutation (a2) in eIF(iso)4E.a, frame-shift mutation (c1) and transposon-insertion mutation (c2). in eIF(iso)4E.c, has been identified respectively from Chinese cabbage. In this study, the differences among three kinds of eIF(iso)4E.a genotype were compared again. Simple and convenient markers were developed. The precise genotyping of eIF(iso)4E.a and eIF(iso)4E.c was detected among 38 hybrids bought from seed market and 96 germplasms created by our breeding team. The results showed that the allele frequencies for A, a1 and a2 were 36.8%, 40.8%, 22.4% among hybrids and 26.1%, 28.1%, 45.8% among germplasms respectively. Total genotype frequency for two mutant alleles of a1 and a2 was 70.1%. Comparatively, only one germplasm carried c1c1 genotype and 13 germplasms carried c2c2 genotype. The total genotype frequency for c1 and c2 was only 10.4%. The results provide molecular markers for utilizing the genes in germplasms detection and new vriaties breeding. They also provide molecular evidence for continuing expanding of those virus-resistance hybrids.
|
|
Received: 30 June 2017
Published: 02 May 2018
|
|
|
|
|
|
| 冯兰香，徐玲，刘佳等.1988. 北京地区大白菜芜菁花叶病毒株系的鉴定[J]. 中国蔬菜,(4):11-13 (Feng L X, Xu L, Liu J et al. 1988. Identification on TuMV strains isolated from Beijing area[J]. China Vegetable, (4):11-13).李国亮.白菜eIF(iso)4E基因对TuMV的抗性机制研究[D].博士学位论文，中国农业科学院，导师：孙日飞. pp.26-32(Li G L.2016. Investigation of interaction mechanisim of the eIF(iso)4E gene to TuMV in Brassica rapa L.[D].Thesis for PHD., Chinese Academy of Agricultural Sciences. Supervisor: Sun R F, pp.26-32)刘栓桃，张志刚，李巧云等. 2014a. 大白菜eIF(iso)4E.a假基因突变体的鉴别及相关检测标记的开发[J].中国农学通报. 2014,30(4):101-107. (Liu S T, Zhang Z G, Li Q Y E et al. 2014a. Characterization of mutation at locus eIF(iso)4E.a and development of related markers in Chinese cabbage[J]. Chines Agricultural Science Bulletin.2014,30(4):101-107.)刘栓桃，张志刚，李巧云等。2014b. 光敏色素B基因(PHYB)启动子突变及与大白菜开花时间的关联分析[J].农业生物技术学报.2014, 22(7): 853~861, doi: 10. 3969/j.issn. 1674- 7968.2014.07.008.(Liu S T, Zhang Z G, Li Q Y E et al. 2014b. Association analysis between phytochrome B gene (PHYB) promoter mutations and flowering time in Chinese cabbage (Brassica rapa L. ssp pekinensis). Journal of Agricultural Biotechnology.2014, 22(7): 853~861, doi: 10. 3969/j.issn. 1674-7968.2014.07.008. )苗立强，张耀伟，崔崇士. 2008. 我国白菜抗病育种研究进展[J]. 东北农业大学学报，37(4): 529-533 (Miao L Q, Zhang Y W, Cui Z S. 2008. The progress of study on Chinese cabbage breeding for diseases resistance in China[J]. Journal of Northeast Agricultural University, 37(4): 529-533).王效睦，白静，谢坤等.2016. 我国大白菜种质资源的研究与利用.山东农业科学. 48(3):143~149. doi: 10. 14083/j.issn.1001-4942.2016.03.035 (Wang X M, Bai J, Xie K et al. 2016. Research and utilization of Chinese cabbage grmplasm resources in China. Shandong Agricultural Sciences. 48(3):143~149.doi: 10. 14083/j.issn.1001-4942.2016.03.035)王雪, 刘玉梅, 李汉霞等. 2005. 芸薹属作物抗芜菁花叶病毒育种研究进展[J].园艺学报，32 (5): 939-946 (Wang X, Liu Y M, Li H X et al. 2005. Advance in research on TuMV-resistance breeding of Brassica crops[J]. Acta Horticulturae Sinica, 32 (5): 939-946.).赵建平, 周钗美, 陈集双等. 2004. 芜菁花叶病毒(TuMV)特性的研究进展[J]. 微生物学通报. 31(6): 100-104 (Zhao J P, Zhou C M, Chen J S et al. 2004. Recent advances in turnip mosaic virus[J]. Microbiology China,31(6): 100-104).Duprat A, Caranta C, Revers F et al.. 2002. The Arabidopsis eukaryotic initiation factor (iso)4E is dispensable for plant growth but required for susceptibility to potyviruses[J]. Plant Journal, 32(6): 927-934. doi: 10.1046/j.1365-313X.2002.01481.x.Jenner C E, Nellist C F, Barker G C, et al. 2010. Turnip mosaic virus (TuMV) is able to use alleles of both eIF4E and eIF(iso)4E from multiple loci of the diploid Brassica rapa[J]. Molecular Plant Microbe Interact, 23(11): 1498-1505. doi: 10.1094/MPMI-05-10-0104.Kim J, Kang W, Yang H et al. 2013. Identification of a broad-spectrum recessive gene in Brassica rapa and molecular analysis of the eIF4E gene family to develop molecular markers[J]. Molecular Breeding, 32(2): 385-398. doi: 10.1007/s11032-013-9878-0.Léonard S, Plante D, Wittmann S et al. 2000. Complex formation between potyvirus VPg and translation eukaryotic initiation factor 4E correlates with virus infectivity[J]. Journal Virology, 74(17): 7730-7737. doi: 10.1128/JVI.74.17.7730-7737.2000.Li Q Y, Zhang X L, Zeng Q et al. 2015. Identification and mapping of a novel turnip mosaic virus resistance gene TuRBCS01 in Chinese cabbage (Brassica rapa L.)[J]. Plant Breeding, 134(2): 221-225. doi: 10.1111/pbr.12239.Liu ST, Li Q Y, Zhang Z G et al., 2017a. Characterization of a Double Mutation of eIF(iso)4E.a and eIF(iso)4E.c with a Diverse Genetic Background and TuMV Resistance in Chinese Cabbage (Brassica rapa L.)[J]. Journal of Agricultural Biotechnology. In publishing.Liu ST, Zhang Z G Wang L H et al., 2017b. Characterization of an active hAT superfamily transposon insertion mutation in eIF(iso)4E[J].c of Chinese cabbage. Journal of Agricultural Biotechnology. In publishing.Nellist C F, Qian W, Jenner C E et al. 2014. Multiple copies of eukaryotic translation initiation factors in Brassica rapa facilitate redundancy, enabling diversification through variation in splicing and broad-spectrum virus resistance[J]. Plant Journal, 77(2): 261-268. doi: 10.1111/tpj.12389.Nicaise V, German-Retana S, Sanjuán R et al. 2003. The eukaryotic translation initiation factor 4E controls lettuce susceptibility to the Potyvirus Lettuce mosaic virus[J]. Plant Physiology, 132(3): 1272-1282. doi: 10.1104/pp.102.017855.Nieto C, Morales M, Orjeda G et al. 2006. An eIF4E allele confers resistance to an uncapped and non-polyadenylated RNA virus in melon[J]. Plant Journal, 48(3): 452-462. doi: 10.1111/j.1365-313X.2006.02885.x.Qian W, Zhang S, Zhang S et al. 2013. Mapping and candidate-gene screening of the novel turnip mosaic virus resistance gene retr02 in Chinese cabbage (Brassica rapa L.) [J]. Theoretical & Applied Genetics, 126(1): 179-188. doi: 10.1007/s00122-012-1972-x.Ruffel S, Dussault M H, Palloix A et al. 2002. A natural recessive resistance gene against potato virus Y in pepper corresponds to the eukaryotic initiation factor 4E (eIF4E)[J]. Plant Journal, 32(6): 1067-1075. doi: 10.1046/j.1365-313X.2002.01499.x.Rusholme R L, Higgins E E, Walsh J A et al. 2007. Genetic control of broad-spectrum resistance to turnip mosaic virus in Brassica rapa(Chinese cabbage) [J]. Journal of General Virology,88(Pt 11): 3177-3186. doi: 10.1099/vir.0.83194-0.Schaad M C, Anderberg R J, Carrington J C. 2000. Strain-specific interaction of the Tobacco etch virus Nia protein with the translation initiation factor eIF4E in the yeast two-hybrid system.Virology, 273(2): 300–306. doi: 10.1006/viro.2000.0416Suh S K, Green S K, Park H G. 1995. Genetics of resistance to five strains of turnip mosaic virus in Chinese cabbage[J]. Euphytica, 81(1):71-77. doi: 10.1007/BF00022460.Walsh J A, Jenner C E. 2002. Turnip mosaic virus and the quest for durable resistance[J]. Molecular Plant Pathology, 3(5):289-300.Wittmann S, Chatel H, Fortin M G, et al. 1997. Interaction of the viral protein genome linked of turnip mosaic potyvirus with the translational eukaryotic initiation factor (iso) 4E of Arabidopsis thaliana using the yeast two-hybrid system[J].Virology, 234(1): 84-92. doi: 10.1006/viro.1997.8634. |
|
|
|
