Abstract
A cytoplasmic male sterile (CMS) line of Brassica juncea was derived by repeated backcrossing of the somatic hybrid (Diplotaxis catholica + B. juncea) to B. juncea. The new CMS line is comparable to euplasmic lines for almost all characters, except for flowers which bear slender, needle-like anthers with aborted pollen. Detailed Southern analysis revealed two copies of coxI gene in the CMS line. One copy, coxI-1 is similar to the coxI gene of B. juncea, whereas the second copy, coxI-2 is present in a novel rearranged region. Northern analysis with eight mitochondrial gene probes showed altered transcript pattern only for the coxI gene. Two transcripts of 2.0 and 2.4 kb, respectively, were detected in the CMS line. The novel 2.4 kb transcript was present in floral bud tissue but absent in the leaf tissue. In plants where male sterility broke down under high temperature during the later part of the growing season, the 2.4 kb coxI transcript was absent, which suggested its association with the CMS. The two coxI genes from the CMS line showed two amino acid changes in the coding region. The novel coxI gene showed unique repeats in the 5′ region suggesting recombination of mitochondrial genomes of the two species. The possible role of the duplicated coxI gene in causing male sterility is discussed.
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References
Abad A. R., Mehrtens B. J. and Mackenzie S. A. 1995 Specific expression in reproductive tissues and fate of a mitochondrial sterility associated protein in cytoplasmic male-sterile bean. Plant Cell 7, 271–285.
Akagi H., Nakamura A., Sawada R., Oka M. and Fiyimura T. 1994 Genetic diagnosis of Cytoplasmic male sterile cybrid plants of rice. Theor. Appl. Genet. 90, 948–951.
Bailey-Serres J., Hanson D. K., Fox T. D. and Leaver C. J. 1986 Mitochondrial genome rearrangement leads to extension and relocation of the cytochrome c oxidase subunit I gene in sorghum. cell 47, 569–576.
Banga S. S., Deol J. S. and Banga S. K. 2003 Alloplasmic male sterile Brassica juncea with Enarthocarpus lyratus cytoplasm and the introgression of gene(s) for fertility restoration from cytoplasm donor species. Theor. Appl. Genet. 106, 1390–1395.
Belliard G., Vedel F. and Pelletier G. 1979 Mitochondrial recombination in cytoplasmic hybrids of Nicotiana tabacum by protoplast fusion. Nature 281, 401–403.
Bonhomme S., Budar F., Lancelin D., Small I., Defrance M. C. and Pelletier G. 1992 Sequence and transcript analysis of the NcoI 2.5 ogura specific fragment correlated with cytoplasmic male sterility in Brassica cybrids. Mol. Gen. Genet. 235, 340–348.
Brown G. G., Formanova N., Jin H., Wargachuk R., Dendy C., Patil P. et al. 2003 The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats. Plant J. 35, 262–272.
Desloire S., Gherbi H., Laloui W., Marhadour S., Clouet V., Cattolico L. et al. 2003 Identification of the fertility restoration locus, Rfo, in radish, as a member of the pentatricopeptide-repeat protein family. EMBO Rep. 4, 588–594.
Forde B. G. and Leaver C. J. 1980 Nuclear and cytoplasmic gene controlling synthesis of variant mitochondrial polypeptides in the male sterile maize. Proc. Natl. Acad. Sci. USA 75, 3841–3845.
Giege P. and Brennicke A. 1999 RNA editing in Arabidopsis mitochondria effects 441 C to U changes in ORFs. Proc. Natl. Acad. Sci. USA 96, 15324–15329.
Grelon M., Budar F., Bonhomme S. and Pelletier G. 1994 Ogura cytoplasmic male-sterility (CMS)-associated orf138 is translated into mitochondrial membrane polypeptide in male-sterile Brassica cybrids. Mol. Gen. Genet. 243, 540–547.
Hanson M. R. and Bentolila S. 2004 Interactions of mitochondrial and nuclear genes that affect male gametophyte development. Plant Cell 16, S154–S169.
Hanson M. R. and Folkerts O. 1992 Structure and function of the higher plant mitochondrial genome. Int. Rev. Cytol. 141, 129–165.
Iwabuchi M., Kyozuka J. and Shimamoto K. 1993. Processing followed by complete editing of an altered mitochondrial atp6 RNA restores fertility of cytoplasmic male sterile rice. EMBO J. 12, 1437–1446.
Iwabuchi M., Koizuka N.; Fujimoto H., Sakai T. and Imamura J. 1999 Identification and expression of the Kosena radish (Raphanus sativus cv Kosena) homologue of the Ogura radish CMS-associated gene, orf138. Plant Mol. Biol. 39, 183–188.
Kempken F. and Pring D. R. 1999 Male sterility in higher plants — fundamentals and applications. Prog. Bot. 60, 140–166.
Kirti P. B., Mohapatra T., Khanna H., Prakash S. and Chopra V. L. 1995a Diplotaxis catholica + Brassica juncea somatic hybrids: molecular and cytogenetic characterization. Plant Cell Rep. 14, 593–597.
Kirti P. B., Banga S. S., Prakash S. and Chopra V. L. 1995b Transfer of Ogu cytoplasmic male sterility to Brassica juncea and improvement of the male sterility through somatic fusion. Theor. Appl. Genet. 91, 517–521.
Koizuka N., Imai R., Fujimoto H., Hayakawa T., Kimura Y., Kohno-Murase J. et al. 2003 Genetic characterization of a pentatricopeptide repeat protein gene, orf 687, that restores fertility in the cytoplasmic male-sterile Kosena radish. Plant J. 34, 407–415.
Krishnasamy S. and Makaroff C. A. 1994 Organ specific reduction in the abundance of a mitochondrial protein accompanies fertility restoration in cytoplasmic male-sterile radish. Plant Mol. Biol. 26, 935–946.
Landgren M., Zetterstrand M., Sundberg E. and Glimelius K. 1996 Alloplasmic male sterile Brassica lines containing B. tournefortii mitochondria express an orf 3′ of the atp6 gene and a 32 kDa protein. Plant Mol. Biol. 32, 870–890.
L’Homme Y. and Brown G. G. 1993 Organisational differences between cytoplasmic male sterile and male fertile Brassica mitochondrial genomes are confined to a single transposed locus. Nucleic Acids Res. 21, 325–335.
L’Homme Y., Stahl R., Li X.-Q., Hameed A. and Brown G. G. 1997 Brassica nap cytoplasmic male sterility is associated with expression of a mtDNA region containing a chimaeric gene similar to the pol CMS-associated orf224 gene. Curr. Genet. 31, 325–335.
Makaroff C. A., Apel I. J. and Palmer J. D. 1989 The atp6 coding region has been disrupted and a novel reading frame generated in the mitochondrial genome of cytoplasmic male-sterile radish. J. Biol. Chem. 264, 11706–11711.
Palmer J. D. 1988 Intraspecific variation and multicircularity in Brassica mitochondrial DNAs. Genetics 118, 341–351.
Pathania A., Bhat S. R., Kumar V. D., Ashutosh, Kirti P. B., Prakash S. and Chopra V. L. 2003 Cytoplasmic male sterility in alloplasmic Brassica juncea carrying Diplotaxis catholica cytoplasm: molecular characterization and genetics of fertility restoration. Theor. Appl. Genet. 107, 455–461.
Pelletier G., Primard C., Ferault M., Vedel F., Chetrit P., Renard M. and Delourme R. 1988 Use of protoplasts in plant breeding: cytoplasmic aspects. Plant Cell Tissue Org. Cult. 12, 173–180.
Prakash S., Kirti P. B., Bhat S. R., Gaikwad K., Kumar V. D. and Chopra V. L. 1998 A Moricandia arvensis based cytoplasmic male sterility and fertility restoration system in Brassica juncea. Theor. Appl. Genet. 97, 488–492.
Prakash S., Ahuja I., Upreti H. C., Kumar D. V., Bhat S. R., Kirti P. B. and Chopra V. L. 2001 Expression of male sterility in alloplasmic Brassica juncea with Erucastrum canariense cytoplasm and the development of fertility restorer system. Plant Breed 120, 1–5.
Pring D. R., Tang H. V., Howad W. and Kempken F. 1999 A unique two-gene gametophytic male sterility system in Sorghum involving a possible role of RNA-editing in fertility restoration. J. Hered. 90, 386–393.
Sambrook J., Fritsch E. F. and Maniatis T. 1989 Molecular cloning: a laboratory manual, 2nd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor.
Schnable P. S. and Wise R. P. 1998 The molecular basis of cytoplasmic male sterility and fertility restoration. Trends Plant Sci. 3, 175–180.
Shinada T., Kikuchi Y., Fujimoto R. and Kishitani S. 2006 An alloplasmic male sterile line of Brassica oleracea harbouring the mitochondria from Diplotaxis muralis expresses a novel chimeric open reading frame orf72. Plant Cell Physiol. 47, 549–553.
Singh M. and Brown G. G. 1991 Suppression of cytoplasmic male sterility by nuclear genes alters expression of a novel mitochondrial gene region. Plant Cell 3, 1349–1362.
Singh M., Hamel N., Menassa R., Li X-Q., Young B., Jean M. et al. 1996 Nuclear genes associated with a single Brassica CMS restorer locus influence transcripts of three mitochondrial gene regions. Genetics 143, 505–516.
Song J. and Hedgecoth C. 1994 A chimeric gene (orf256) is expressed as protein only in cytoplasmic male sterile lines of wheat. Plant Mol. Biol. 26, 535–539.
Tang H. V., Chang R. and Pring D. R. 1998 Cosegregation of single genes associated with fertility restoration and transcript processing of Sorghum mitochondrial orf107 and urf209. Genetics 150, 383–391.
Zabala G., Gabay-Laughnan S. and Laughnan J. R. 1997 The nuclear gene RF3 affects the expression of the mitochondrial chimeric sequence R implicated in S-type male sterility in maize. Genetics 147, 847–860.
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Pathania, A., Kumar, R., Kumar, V.D. et al. A duplicated coxI gene is associated with cytoplasmic male sterility in an alloplasmic Brassica juncea line derived from somatic hybridization with Diplotaxis catholica . J Genet 86, 93–101 (2007). https://doi.org/10.1007/s12041-007-0014-y
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DOI: https://doi.org/10.1007/s12041-007-0014-y