Abstract
BackgroundThe application of hybrid rice technology has significantly increased global rice production during the last three decades. Approximately 90% of the commercially cultivated rice hybrids have been derived through three-line breeding involving the use of WA-CMS lines. It is believed that during the 21st century, hybrid rice technology will make significant contributions to ensure global food security. This study examined the poorly understood molecular basis of the WA-CMS system in rice.ResultsRFLPs were detected for atp6 and orfB genes in sterile and fertile rice lines, with one copy of each in the mt-genome. The RNA profile was identical in both lines for atp6, but an additional longer orfB transcript was identified in sterile lines. 5' RACE analysis of the long orfB transcript revealed it was 370 bp longer than the normal transcript, with no indication it was chimeric when compared to the genomic DNA sequence. cDNA clones of the longer orfB transcript in sterile lines were sequenced and the transcript was determined unedited. Sterile lines were crossed with the restorer and maintainer lines, and fertile and sterile F1 hybrids were respectively generated. Both hybrids contained two types of orfB transcripts. However, the long transcript underwent editing in the fertile F1 hybrids and remained unedited in the sterile lines. Additionally, the editing of the 1.1 kb orfB transcript co-segregated with fertility restoring alleles in a segregating population of F2 progeny; and the presence of unedited long orfB transcripts was detected in the sterile plants from the F2 segregating population.ConclusionThis study helped to assign plausible operative factors responsible for male-sterility in the WA cytoplasm of rice. A new point of departure to dissect the mechanisms governing the CMS-WA system in rice has been identified, which can be applied to further harness the opportunities afforded by hybrid vigor in rice.
Highlights
The application of hybrid rice technology has significantly increased global rice production during the last three decades
In almost all plant Cytoplasmic male sterility (CMS) systems studied to date, the male sterility trait was associated with changes in mitochondrial gene organization. [4] demonstrated that cytoplasmic male sterility was caused by protein defects involved in mitochondrial energy production and often involved ATP synthase subunit genes
Structural organization of atp6, atpA, atp9 and orfB in sterile and fertile rice lines The organization of four mitochondrion-encoded genes was examined by Southern blot analysis of the CMS rice line APMS-6A, including the corresponding maintainer APMS-6B and restorer BR-1870 lines
Summary
The application of hybrid rice technology has significantly increased global rice production during the last three decades. In almost all plant CMS systems studied to date, the male sterility trait was associated with changes in mitochondrial gene organization. [4] demonstrated that cytoplasmic male sterility was caused by protein defects involved in mitochondrial energy production and often involved ATP synthase subunit genes. Mt-DNA rearrangement has been shown to generate novel chimeric ORFs, which resulted in the expression of novel polypeptides [5]. Impairment of functions of mitochondrial genes have been shown to be associated with CMS [4,5,11,12]. The precise relationship between mitochondrial CMS-associated genes and male sterility varies from species to species and is poorly understood
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