Mitochondrial-targeted Expression of orf456 Causes Male Sterility in Chinese Cabbage (Brassica rapa L.)

Cytoplasmic male sterility (CMS) is a phenomenon specific to plant reproduction that has been extensively exploited for hybrid seed production. Orf456 was previously identified as a candidate gene mediating male sterility in pepper. To gain further insight of this candidate gene, we carried out experiments to transform Chinese cabbage (Brassica rapa L.). About 30 T1 transgenic lines were obtained and approximately 50 % of T1 transgenic Chinese cabbage lines showed male-sterility. To evaluate pollen viability, three different approaches including plasmolysis test, staining pollen and in vitro germination assay were used. Analysis of the CMS transgenic lines showed that trasgenic Chinese cabbages produced aberrant pollen development while some were unable to produce pollen. In conclusion, the mitochondrial orf456 gene could induce partial male sterility in transgenic Chinese cabbage. Nevertheless, how the orf456 gene precisely functions to induce male sterility and its biochemical function remains to be discovered.

[1]  Tom Lawrenson,et al.  Brassica rapa. , 2015, Methods in molecular biology.

[2]  N. Ramchiary,et al.  Quantitative Trait Loci Mapping in Brassica rapa Revealed the Structural and Functional Conservation of Genetic Loci Governing Morphological and Yield Component Traits in the A, B, and C Subgenomes of Brassica Species , 2012, DNA research : an international journal for rapid publication of reports on genes and genomes.

[3]  S. Ahmad,et al.  Expression of Heterosis for Productive Traits in F 1 Brinjal ( Solanum melongena L.) Hybrids , 2011 .

[4]  Mingfang Zhang,et al.  Mitochondrially-targeted expression of a cytoplasmic male sterility-associated orf220 gene causes male sterility in Brassica juncea , 2010, BMC Plant Biology.

[5]  Jianbo Wang,et al.  A comparative light and electron microscopic analysis of microspore and tapetum development in fertile and cytoplasmic male sterile radish , 2010, Protoplasma.

[6]  Mingfang Zhang,et al.  Molecular Genetics of Mitochondrial Respiratory / ATP-Related Genes in Relation to Cytoplasmic Male-Sterility of Higher Plants , 2009 .

[7]  Y. Onodera,et al.  A male sterility-associated mitochondrial protein in wild beets causes pollen disruption in transgenic plants. , 2008, The Plant journal : for cell and molecular biology.

[8]  C. Chase Cytoplasmic male sterility: a window to the world of plant mitochondrial-nuclear interactions. , 2007, Trends in genetics : TIG.

[9]  Byung-Dong Kim,et al.  Isolation and characterization of the cytoplasmic male sterility-associated orf456 gene of chili pepper (Capsicum annuum L.) , 2007, Plant Molecular Biology.

[10]  T. Börner,et al.  Mitochondrial effects on flower and pollen development. , 2005, Mitochondrion.

[11]  Byung-Dong Kim,et al.  The organization of mitochondrial atp6 gene region in male fertile and CMS lines of pepper (Capsicum annuum L.) , 2005, Current Genetics.

[12]  S. Bentolila,et al.  Interactions of Mitochondrial and Nuclear Genes That Affect Male Gametophyte Development , 2004, The Plant Cell Online.

[13]  J. Edqvist,et al.  Male-sterile tobacco displays abnormal mitochondrial atp1 transcript accumulation and reduced floral ATP/ADP ratio , 2000, Plant Molecular Biology.

[14]  J. Balk,et al.  ORFB is a subunit of F1 FO‐ATP synthase: insight into the basis of cytoplasmic male sterility in sunflower , 2003, EMBO reports.

[15]  Byung-Dong Kim,et al.  Identification of coxII and atp6 Regions as Associated to CMS in Capsicum annuum by Using RFLP and Long Accurate PCR , 2001 .

[16]  N. Nassar,et al.  Brief report : the transference of apomixis genes from manihot neusana nassar to cassava, M. esculenta crantz , 2004 .

[17]  M. Tuinstra,et al.  Estimation of pollen viability in grain sorghum. , 2000 .

[18]  C. Gómez-Campo Biology of brassica coenospecies , 1999 .

[19]  P. Schnable,et al.  The molecular basis of cytoplasmic male sterility and fertility restoration , 1998 .

[20]  J. Prince,et al.  A versatile and high-yielding protocol for the preparation of genomic DNA from Capsicum spp. (pepper) , 1997 .

[21]  A. Abad,et al.  A cytoplasmic male sterility-associated mitochondrial protein causes pollen disruption in transgenic tobacco. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[22]  C. S. Levings,et al.  URF13, a ligand-gated, pore-forming receptor for T-toxin in the inner membrane ofcms-T mitochondria , 1995, Journal of bioenergetics and biomembranes.

[23]  H. Ogura 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 , 1967 .

[24]  A. Nassar,et al.  The transference of apomixis genes from Manihot neusana Nassar to cassava, M. esculenta Crantz By , 2022 .