Leaf Morphology and Ultrastructure Responses to Elevated O3 in Transgenic Bt (cry1Ab/cry1Ac) Rice and Conventional Rice under Fully Open-Air Field Conditions

Background: Elevated tropospheric ozone severely affects not only yield but also the morphology, structure and physiological functions of plants. Because of concerns regarding the potential environmental risk of transgenic crops, it is important to monitor changes in transgenic insect-resistant rice under the projected high tropospheric ozone before its commercial release. Methodology/Principal Findings: Using a free-air concentration enrichment (FACE) system, we investigated the changes in leaf morphology and leaf ultrastructure of two rice varieties grown in plastic pots, transgenic Bt Shanyou 63 (Bt-SY63, carrying a fusion gene of cry1Ab and cry1Ac) and its non-transgenic counterpart (SY63), in elevated O-3 (E-O-3) versus ambient O-3 (A-O-3) after 64-DAS (Days after seeding), 85-DAS and 102-DAS. Our results indicated that E-O-3 had no significant effects on leaf length, leaf width, leaf area, stomatal length and stomatal density for both Bt-SY63 and SY63. E-O-3 increased the leaf thickness of Bt-SY63, but decreased that of SY63. O-3 stress caused early swelling of the thylakoids of chloroplasts, a significant increase in the proportion of total plastoglobule area in the entire cell area (PCAP) and a significant decrease in the proportion of total starch grain area in the entire cell area (SCAP), suggesting that E-O-3 accelerated the leaf senescence of the two rice genotypes. Compared with SY63, E-O-3 caused early swelling of the thylakoids of chloroplasts and more substantial breakdown of chloroplasts in Bt-SY63. Conclusions/Significance: Our results suggest that the incorporation of cry1Ab/Ac into SY63 could induce unintentional changes in some parts of plant morphology and that O-3 stress results in greater leaf damage to Bt-SY63 than to SY63, with the former coupled with higher O-3 sensitivity in CCAP (the proportions of total chloroplast area in the entire cell area), PCAP and SCAP. This study provides valuable baseline information for the prospective commercial release of transgenic crops under the projected future climate.