BACKGROUND: Frequent fungal diseases tend to lead to severe losses in rice production. As a main component of the fungal cell wall, glucan plays an important role in the growth and development of fungi. Glucanase can inhibit the growth of fungi by breaking glycosidic bonds, and may be a promising target for developing rice varieties with broad-spectrum disease resistance.RESULTS: We transferred a codon-optimized ⊎-1,6-glucanase gene (GluM) from myxobacteria into the japonica rice variety Zhonghua11 (ZH11), and obtained a large number of individual transgenic plants with GluM overexpression. Based on molecular analysis, three single-copy homozygous lines with GluM overexpression were selected for assessment of fungal disease resistance at the T 3 generation. Compared with that of the recipient cultivar ZH11, the area of rice blast lesion in transgenic rice was reduced by 82.71%; that of sheath blight lesion was decreased by 35.76%-43.67%; the sheath blight resistance in the field was enhanced by an average of 0.75 grade over 3 years; and the incidence of diseased panicles due to rice false smut was decreased by 65.79%. More importantly, there was no obvious loss of yield (without a significant effect on agronomic traits). Furthermore, plants overexpressing a ⊎-1,6-glucanase gene showed higher disease resistance than rice plants overexpressing a ⊎-1,3-glucanase gene derived from tobacco.CONCLUSION: The ⊎-1,6-glucanase gene GluM can confer broad-spectrum disease resistance to rice, providing an environmentally friendly alternative way to effectively manage fungal pathogens in rice production.

ABSTRACT. Ultrasound-targeted microbubble destruction (UTMD) has been recently developed for destroying bubbles carrying drugs or genes, thereby permitting local release of these target molecules. We investigated whether SonoVue ® , a new contrast agent that contains phospholipid-stabilized microbubbles filled with sulfur hexafluoride vapor, is effective at delivering a recombinant adeno-associated viral (rAAV) vector to the rat heart by UTMD. Serotype-2 (rAAV2) marked with green fluorescent protein (GFP) as a reporter gene was attached to the surface of sulfur hexafluoride-filled microbubbles. Microbubbles were infused into the tail vein of rats with or without simultaneous echocardiography. Additional controls included ultrasound microbubbles that did not contain virus, virus alone, and virus plus ultrasound. One group underwent echocardiographic destruction of microbubbles followed by rAAV2-GFP infusion. Rats were killed after 4 weeks and examined for GFP expression. Green fluorescence was detected in all groups that received the rAAV2-GFP vector, indicating expression of the rAAV2 1319 ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 12 (2): 1318-1326 (2013 Recombinant adeno-associated virus transgene transgene; however, GFP expression in the UTMD group was significantly higher than that in control groups. We conclude that ultrasoundmediated destruction mediated by SonoVue is a promising method for delivery of rAAV2 to the heart in vivo.

BACKGROUND The striped stem borer (SSB), Chilo suppressalis Walker, is a major pest of rice worldwide. Breeding of transgenic rice expressing Bacillus thuringiensis (Bt) toxins is a powerful strategy to control SSB. However, pests may evolve certain resistance to Bt toxins in transgenic plants. Hence, new controlling strategies must be continuously developed. RESULTS We successfully generated SSB‐resistant rice (csu‐53) expressing the artificial microRNA (amiRNA) of SSB endogenous miRNA (csu‐novel‐miR53) through the RNAi‐based technology. Feeding assays demonstrated that csu‐53 rice inhibited larval growth, delayed pupation time, and reduced pupal weight and eclosion rate of SSB larva. In a 10‐day feeding experiment, the miRNA mimic of csu‐novel‐miR53 also suppressed larval growth and more importantly increased larval mortality. Transcriptome analysis identified 28 differentially expressed unigenes (DEGs) in the midgut between SSB larvae fed on csu‐53 rice and the wild type. One DEG (DN90065_c0_g12) validated by qRT‐PCR had a predicted target site of csu‐novel‐miR53. In addition, in vitro double‐stranded RNA synthesis and further feeding assay proved that DN90065_c0_g12 is most likely the target of csu‐novel‐miR53. CONCLUSION amiRNA‐mediated strategy can be applied to the development of insect‐resistant crops, and the novel amiRNA csu‐novel‐miR53 of SSB has important application potential in developing SSB resistant rice. © 2021 Society of Chemical Industry.