Gene-Based Marker to Differentiate Among B, A, and R Lines in Hybrid Production of Rapeseed Ogura System

Document Type: Research Paper

Authors

1 Department of Biotechnology, Faculty of Agriculture, Tarbiat Modarres University, Tehran, Iran

2 Seed and Plant Improvement Institute (SPII), Agricultural Research, Education and Extension (AREEO), Shahid Fahmideh BLVRD, Seed and Plant Improvement Institute Campus, Karaj, Iran

3 Department of Plant Genetics and Breeding, Faculty of Agriculture, Tarbiat Modarres University, Tehran, Iran

Abstract

Background: In plant breeding program to produce hybrid varieties, pair of male sterile and restorer fertility lines are required. Differentiation of lines possessing restorer fertility allele from the lines lacking it remove the need for the progeny test, and thus reducing the time and the cost in the hybrid production procedure. Canola breeding program in Iran has concentrated toward production of domestic hybrid varieties, however, it suffers from lack of molecular information in restore fertility status of lines, and therefore it needs time and tedious activities.
Objectives: To design gene-based markers for distinguishing R-, A- lines and hybrids in sunflower breeding programs.
Material and Methods: Aligning sequences of locus responsible for male sterility and that of male fertility resulted in finding differences in the loci, which used to define two set of suitable primer pairs. Genomic DNA from 25 R-lines (23 inbred lines and two commercial lines), 9 A-lines (7 inbred lines and two commercial lines), one B-line and two commercial hybrids were extracted and used in PCR as template.
Results: Using one-primer pairs, a band of nearly 1500 bp was amplified in restorer lines but not in A-, B- lines. Another primer pair used to distinguish hybrids (heterozygout) from restorer lines. Results of the report is predicted to be used in canola breeding for hybrid production.
Conclusions: Although the molecular bases for the male sterility and fertility restoration in rapeseed is not published, taking advantages of gene-based markers, make rapeseed breeding program more efficient regarding time and costs.

Keywords

Main Subjects


1.           Burns DR, Forhan MA, Barnes S, Buzza GC, Stoenescu FM, Huskowska T. Cytoplasmic male sterility system production canola hybrids. Google Patents; 2001.

2.           Oghan H. Canola hybrid production in Iran. Najafabadi M, editor. Karaj2017.

3.           Chen L, Liu YG. Male sterility and fertility restoration in crops. Annu Rev Plant Biol. 2014;65:579-606. doi: 10.1146/annurev-arplant-050213-040119 pmid: 24313845

4.           Touzet P, Meyer EH. Cytoplasmic male sterility and mitochondrial metabolism in plants. Mitochondrion. 2014;19 Pt B:166-171. doi: 10.1016/j.mito.2014.04.009 pmid: 24769053

5.           Brown GG, Formanova N, Jin H, Wargachuk R, Dendy C, Patil P, et al. The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats. Plant J. 2003;35(2):262-272. doi: 10.1046/j.1365-313x.2003.01799.x pmid: 12848830

6.           Singh M, Brown GG. Suppression of cytoplasmic male sterility by nuclear genes alters expression of a novel mitochondrial gene region. Plant Cell. 1991;3(12):1349-1362. doi: 10.1105/tpc.3.12.1349 pmid: 1840901

7.           Brown GG. Unique aspects of cytoplasmic male sterility and fertility restoration in Brassica napus. Journal of Heredity. 1999;90(3):351-356. doi: 10.1093/jhered/90.3.351

8.           Uyttewaal M, Arnal N, Quadrado M, Martin-Canadell A, Vrielynck N, Hiard S, et al. Characterization of Raphanus sativus pentatricopeptide repeat proteins encoded by the fertility restorer locus for Ogura cytoplasmic male sterility. Plant Cell. 2008;20(12):3331-3345. doi: 10.1105/tpc.107.057208 pmid: 19098270

9.           Ogura H. Studies on the new male-sterility in Japanese radish, with special reference to the utilization of this sterility towerds the practical raising of hybrid seeds. Memoirs of the Faculty of Agriculture, Kagoshima University: Kagoshima University; 1968. p. 39-78.

10.        Desloire S, Gherbi H, Laloui W, Marhadour S, Clouet V, Cattolico L, et al. Identification of the fertility restoration locus, Rfo, in radish, as a member of the pentatricopeptide-repeat protein family. EMBO Rep. 2003;4(6):588-594. doi: 10.1038/sj.embor.embor848 pmid: 12740605

11.        Mikami T. [Molecular basis of cytoplasmic male sterility in higher plants]. Tanpakushitsu Kakusan Koso. 1992;37(7):1130-1136. pmid: 1603956

12.        Sjolander A, Hossjer O, Hartman LW, Humphreys K. Fine mapping of disease genes using tagging SNPs. Ann Hum Genet. 2007;71(Pt 6):815-827. doi: 10.1111/j.1469-1809.2007.00379.x pmid: 17587339

13.        Khlestkina EK, Pestsova EG, Salina E, Roder MS, Arbuzova VS, Koval SF, et al. Genetic mapping and tagging of wheat genes using RAPD, STS and SSR markers. Cell Mol Biol Lett. 2002;7(2B):795-802. pmid: 12378240

14.        Sharma RP, Mohapatra T. Molecular mapping and tagging of genes in crop plants. Genetica. 1996;97(3):313-320. doi: 10.1007/bf00055317 pmid: 9081859

15.        Nayak SN, Zhu H, Varghese N, Datta S, Choi HK, Horres R, et al. Integration of novel SSR and gene-based SNP marker loci in the chickpea genetic map and establishment of new anchor points with Medicago truncatula genome. Theor Appl Genet. 2010;120(7):1415-1441. doi: 10.1007/s00122-010-1265-1 pmid: 20098978

16.        Chin JH, Lu X, Haefele SM, Gamuyao R, Ismail A, Wissuwa M, et al. Development and application of gene-based markers for the major rice QTL Phosphorus uptake 1. Theor Appl Genet. 2010;120(6):1073-1086. doi: 10.1007/s00122-009-1235-7 pmid: 20035315

17.        Murray MG, Thompson WF. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980;8(19):4321-4325. doi: 10.1093/nar/8.19.4321 pmid: 7433111

18.        Fakharian M. Parents for canola hybrids. Najafabadi M, editor. Karaj2017.

19.        Hernandez Mora JR, Rivals E, Mireau H, Budar F. Sequence analysis of two alleles reveals that intra-and intergenic recombination played a role in the evolution of the radish fertility restorer (Rfo). BMC Plant Biol. 2010;10:35. doi: 10.1186/1471-2229-10-35 pmid: 20178653