An unedited 1.1 kb mitochondrial orfB gene transcript in the Wild Abortive Cytoplasmic Male Sterility (WA-CMS) system of Oryza sativa L. subsp. indica

<p>Abstract</p> <p>Background</p> <p>The 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 21^stcentury, 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.</p> <p>Results</p> <p>RFLPs were detected for <it>atp6 </it>and <it>orfB </it>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 <it>atp6</it>, but an additional longer <it>orfB </it>transcript was identified in sterile lines. 5' RACE analysis of the long <it>orfB </it>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 <it>orfB </it>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 F₁hybrids were respectively generated. Both hybrids contained two types of <it>orfB </it>transcripts. However, the long transcript underwent editing in the fertile F₁hybrids and remained unedited in the sterile lines. Additionally, the editing of the 1.1 kb <it>orfB </it>transcript co-segregated with fertility restoring alleles in a segregating population of F₂progeny; and the presence of unedited long <it>orfB </it>transcripts was detected in the sterile plants from the F₂segregating population.</p> <p>Conclusion</p> <p>This 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.</p

Prediction-based protein engineering of domain I of Cry2A entomocidal toxin of Bacillus thuringiensis for the enhancement of toxicity against lepidopteran insects

Issues relating to sustenance of the usefulness of genetically modified first generation Bt crop plants in the farmer’s field are of great concern for crop scientists. Additional biotechnological strategies need to be in place to safeguard the possibility for yield loss of Bt crop by other lepidopteran insects that are insensitive to the Cry1A toxin, and also against the possibility for emergence of resistant insects. In this respect, Cry2A toxin has figured as a prospective candidate to be the second toxin to offer the required protection along with Cry1A. In the present study, the entomocidal potency of Cry2A toxin was enhanced through knowledge-based protein engineering of the toxin molecule. Deletion of 42 amino acid residues from the N-terminal end of the peptide followed by the replacement of Lys residues by nonpolar amino acids in the putative transmembrane region including the introduction of Pro resulted in a 4.1–6.6-fold increase in the toxicity of the peptide against three major lepidopteran insect pests of crop plants