Ethylene insensitive 2 (EIN2) as a potential target gene to enhance Fusarium head blight disease resistance.

Fusarium head blight (FHB) caused by Fusarium graminearum (Fg) severely affects cereal crops, especially wheat and barley. FHB results in significant yield loss, reduces grain quality and contaminates grains with mycotoxin. The development of FHB-resistant cereal cultivars can be expedited through CRISPR gene editing. The Arabidopsis ethylene insensitive 2 (AtEIN2) plays a key role in ethylene signaling pathway and is critical for monitoring plant growth and defense responses. RNAi down-regulation of the wheat homolog TaEIN2 has been shown to enhance wheat FHB resistance. Here we generated site-specific mutations in AtEIN2 by CRISPR-editing. Detached inflorescence infection assays revealed that AtEIN2 knock-out (KO) mutants displayed enhanced Fg resistance and substantially reduced Fg spore production in planta. Gene expression profiling of defense genes revealed that impairment of AtEIN2 resulted in down-regulation of the ethylene signaling pathway while the salicylic acid signaling pathway was unaffected. Complementation of AtEIN2-KO plants with a barley orthologue, HvEIN2, restored Fg susceptibility, indicating that HvEIN2 is functionally equivalent to its Arabidopsis counterpart and, hence, may have a similar role in conditioning barley Fg susceptibility. These results provide insight into the defense role of EIN2 and a molecular and functional foundation for manipulating HvEIN2 to enhance FHB resistance in barley.

CRISPR-Editing of Sweet Basil (Ocimum basilicum L.) Homoserine Kinase Gene for Improved Downy Mildew Disease Resistance.

Sweet basil (Ocimum basilicum L.) downy mildew disease (DM) caused by Peronospora belbahrii is a worldwide threat to the basil industry due to the lack of natural genetic resistance in sweet basil germplasm collections. In this study, we used CRISPR-gene editing to modify the sweet basil DM susceptibility gene homoserine kinase (ObHSK). Gene-edited plants challenged with P. belbahrii displayed a significantly reduced susceptibility to DM, based on phenotypic disease indices and on in planta pathogen load. These results suggest that ObHSK plays a role in conditioning DM susceptibility, similar to that observed for the AtHSK gene in Arabidopsis. These results demonstrate the utility of CRISPR-gene editing in enhancing DM resistance and contributing to sweet basil breeding programs.

Validation of barley 2OGO gene as a functional orthologue of Arabidopsis DMR6 gene in Fusarium head blight susceptibility.

Fusarium head blight (FHB) caused by Fusarium graminearum (Fg) is a devastating disease of crops, especially wheat and barley, resulting in significant yield loss and reduced grain quality. Fg infection leads to the production of mycotoxins, whose consumption is toxic to humans and livestock. The Arabidopsis DMR6 gene encodes a putative 2-oxoglutarate Fe(II)-dependent oxygenase (2OGO) and has been identified as a susceptibility factor to downy mildew. We generated site-specific mutations in Arabidopsis At2OGO by CRISPR/Cas9 gene editing. The resulting At2OGO knock-out (KO) mutants display enhanced resistance to Fg in a detached inflorescence infection assay. Expression profiling of defense genes revealed that impairment of At2OGO function resulted in the upregulation of defense genes that are regulated by salicylic acid (SA), jasmonic acid (JA) and ethylene (ET) pathways. Complementation of the At2OGO-KO lines with a barley (cv. Conlon) orthologue, Hv2OGO, restored susceptibility to Fg. This result indicates that the Hv2OGO gene is functionally equivalent to its Arabidopsis counterpart and, hence, may have a similar role in conditioning susceptibility to FHB in barley. These results provide a molecular basis for proposing 2OGO as a plant immunity suppressor in Arabidopsis and potentially in barley plants and establish a rationale and strategy for enhancing FHB resistance in barley.

Draft Genome Sequence of Streptomyces aureoverticillatus HN6, a Strain Antagonistic against Fusarium oxysporum f. sp. cubense Race 4.

Streptomyces aureoverticillatus HN6 was isolated in Hainan, China. It is highly inhibitory to Fusarium oxysporum f. sp. cubense race 4 (FOC4), which causes banana Fusarium wilt. The HN6 genome was fully sequenced and assembled. Bionformatic analysis shows that the HN6 genome contains at least 208 genes involved in antibiotic biosynthesis.