WUSCHEL-related homeobox (WOX) transcription factors (TFs) are well known for their role in plant development but are rarely studied in citrus. In this study, we identified 11 putative genes from the sweet orange genome and divided the citrus WOX genes into three clades (modern/WUSCHEL(WUS), intermediate, and ancient). Subsequently, we performed syntenic relationship, intron-exon organization, motif composition, and cis-element analysis. Co-expression analysis based on RNA-seq and tissue-specific expression patterns revealed that CsWOX gene expression has multiple intrinsic functions. CsWUS homolog of AtWUS functions as a transcriptional activator and binds to specific DNA. Overexpression of CsWUS in tobacco revealed dramatic phenotypic changes, including malformed leaves and reduced gynoecia with no seed development. Silencing of CsWUS in lemon using the virus-induced gene silencing (VIGS) system implied the involvement of CsWUS in cells of the plant stem. In addition, CsWUS was found to interact with CsCYCD3, an ortholog in Arabidopsis (AtCYCD3,1). Yeast one-hybrid screening and dual luciferase activity revealed that two TFs (CsRAP2.12 and CsHB22) bind to the promoter of CsWUS and regulate its expression. Altogether, these results extend our knowledge of the WOX gene family along with CsWUS function and provide valuable findings for future study on development regulation and comprehensive data of WOX members in citrus.
Commercially important palms (oil palm, coconut, and date palm) are widely grown perennial trees with tremendous commercial significance due to food, edible oil, and industrial applications. The mounting pressure on the human population further reinforces palms’ importance, as they are essential crops to meet vegetable oil needs around the globe. Various conventional breeding methods are used for the genetic improvement of palms. However, adopting new technologies is crucial to accelerate breeding and satisfy the expanding population’s demands. CRISPR/Cas9 is an efficient genome editing tool that can incorporate desired traits into the existing DNA of the plant without losing common traits. Recent progress in genome editing in oil palm, coconut and date palm are preliminarily introduced to potential readers. Furthermore, detailed information on available CRISPR-based genome editing and genetic transformation methods are summarized for researchers. We shed light on the possibilities of genome editing in palm crops, especially on the modification of fatty acid biosynthesis in oil palm. Moreover, the limitations in genome editing, including inadequate target gene screening due to genome complexities and low efficiency of genetic transformation, are also highlighted. The prospects of CRISPR/Cas9-based gene editing in commercial palms to improve sustainable production are also addressed in this review paper.
Oil palm (Elaeis guineensis) is the highest oil-yielding commercially grown perennial tree. Oil palm germplasm conservation and in vitro clonal propagation strengthened the world’s efforts to ensure future food security. Cryopreservation provides long-term storage for germplasm. The storage of plant material at cryogenic temperatures (−196 °C) following dehydration causes cryoinjury. The cryotolerance mechanism has rarely been studied in oil palm zygotic embryos (ZE) and embryogenic calli (EC). A simple and effective cryopreservation method was established for ZE. ZE surrounded by endosperm was air-dried for 3 days without any complicated chemical pre-treatments before cryopreservation, while the viability rate and following germination rate could reach up to 96.67% and 90.88%, respectively. As for EC, the preferred method could be pre-culture in liquid MS medium with 0.3 M sucrose for 12 h and PVS2 treatment for 5 min prior to cryopreservation, and the viability rate reached 68.33%. SSR markers were used to verify the genetic stability after cryopreservation. In addition, changes in enzyme activities (CAT, POD, and SOD) showed a consistent trend with H2O2 production among ZE samples, indicating that these antioxidants were involved in ROS scavenging. Furthermore, differently expressed genes (DEGs) related to ROS, osmotic, and cold stress responses were selected for correlation network analysis. Most genes involved in ROS production (RBOH, PAO, and PRX) and ROS scavenging (APX, PER, SOD, CAT, GPX, and AOX) showed higher expression levels in EC, suggesting that EC was more sensitive to oxidative stress than ZE. The cryotolerance mechanism was further summarized accordingly. These results contributed to cryopreservation methods and provided a better understanding of cryotolerance in oil palm.
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