Huanglongbing (HLB) is one of the most severe citrus diseases in the world, causing huge economic losses. However, efficient methods of protecting citrus from HLB have not yet been developed. microRNA (miRNA)-mediated regulation of gene expression is a useful tool to control plant diseases, but the miRNAs involved in regulating resistance to HLB have not yet been identified. In this study, we found that miR171b positively regulated resistance to HLB in citrus. Upon infection with HLB bacteria, the bacteria were detected in the second month in the control plants. However, in the miR171b-overexpressing transgenic citrus plants, the bacteria could not be detected until the 24th month. RNA-seq data indicated that multiple pathways, such as photosynthesis, plant–pathogen interaction, the MAPK signaling pathway, etc., might be involved in improving the resistance to HLB in miR171b-overexpressing plants compared with the control. Finally, we determined that miR171b could target SCARECROW-like (SCL) genes to downregulate their expression, which then led to promoted resistance to HLB stress. Collectively, our results demonstrate that miR171b plays a positive regulatory role in resistance to citrus HLB, and provides a new insight into the role of miRNAs in the adaptation of citrus to HLB stress.

Previous studies have shown that abscisic acid and ethylene are involved in pulp maturation and peel coloration in the nonclimacteric citrus fruits. There are also signs indicating that other plant hormones may play some roles in citrus fruit ripening. In this study, we compared profiles of genome-wide gene expression, and changes in hormones and peel pigments between fruits of Shatangju mandarin (Citrus reticulata Blanco, designated WT) and its natural mutant, Yuenongwanju (designated MT). The MT fruit mature approximately two months later than the WT fruit. Significant differences in fruit diameter, total soluble solids, titratable acid content, chlorophylls and carotenoids were detected between the fruits of the two genotypes at the sampled time points. Genome-wide transcriptome profiling showed that many genes involved in auxin and ABA metabolism and/or signaling pathways were differentially expressed between the MT and the WT fruits. Importantly, the expression of CrYUCCA8 was significantly lower and the expression of CrNCED5 was significantly higher in WT than in MT fruits at 230 and 250 DPA, respectively. In addition, the IAA level in the MT fruit was significantly higher than that in the WT counterpart, whereas a significantly lower level of ABA was detected in the mutant. Treatment of the WT fruit with exogenous IAA significantly delayed fruit maturation. Our results provide experimental evidence supporting the notion that auxin is a negative regulator of fruit maturation in citrus.

The study focuses on the Cyclic nucleotide-gated ion channels (CNGCs) proteins in citrus, aiming to investigate their potential roles. A total of 33 CcCNGC proteins were identified and characterized in Citrus clementina using a genome-wide method. The study revealed that these proteins share a conserved CNGC domain structurally but exhibit significant differences in their primary sequence and motif composition. Phylogenetic analysis classified the CcCNGC proteins into 13 subgroups. The cis-elements present in all CcCNGCs promoters were identified and classified, and the number of elements was determined. The results suggested that these genes play important roles in citrus growth and development, as well as in response to biotic and abiotic stresses. Gene expression analysis further supported these findings, demonstrating that CNGC genes were responsive to various plant hormones and Phytophthora nicotianae infection, which causes citrus foot rot. Overall, the study indicated that members of the CcCNGC gene family exhibit structural and functional diversity. Further research is needed to validate the specific functions of individual family members and their roles in citrus physiology and response to stress conditions.

The Dof transcription factor family plays crucial roles in plant growth and stress responses. In this study, we identified 24 Dof genes (CcDof1–CcDof24) from the genome of Citrus clementina (Hort. ex Tan.). Phylogenetic analysis classified these proteins into six distinct clades, revealing evolutionary conservation with Dof members from Arabidopsis and tomato. Analysis of gene structure and conserved motifs showed that most CcDof genes are intronless or contain only a few introns, and their motif compositions are largely consistent with their phylogenetic relationships. Promoter analysis revealed a variety of cis-regulatory elements associated with light responsiveness, hormone signaling, and abiotic/biotic stress responses. Expression profiling demonstrated that CcDof genes exhibit tissue-specific expression patterns and are differentially regulated by various phytohormones (including ABA, SA, GA, and MeJA), low temperature stress, and infection by Phytophthora parasitica. Notably, transient overexpression of CcDof4 and CcDof6 in citrus leaves significantly enhanced resistance to P. parasitica, accompanied by upregulation of SA pathway markers NPR1 and PR1. Our findings provide a systematic characterization of the CcDof family and highlight the important roles of CcDof4 and CcDof6 in mediating citrus disease resistance, likely through modulation of the SA signaling pathway.