Worldwide, citrus is one of the most important fruit crops and is grown in more than 130 countries, predominantly in tropical and subtropical areas. The healthy progress of the citrus industry has been seriously affected by biotic and abiotic stresses. Several diseases, such as canker and huanglongbing, etc., rigorously affect citrus plant growth, fruit quality, and yield. Genetic engineering technologies, such as genetic transformation and genome editing, represent successful and attractive approaches for developing disease-resistant crops. These genetic engineering technologies have been widely used to develop citrus disease-resistant varieties against canker, huanglongbing, and many other fungal and viral diseases. Recently, clustered regularly interspaced short palindromic repeats (CRISPR)-based systems have made genome editing an indispensable genetic manipulation tool that has been applied to many crops, including citrus. The improved CRISPR systems, such as CRISPR/CRISPR-associated protein (Cas)9 and CRISPR/Cpf1 systems, can provide a promising new corridor for generating citrus varieties that are resistant to different pathogens. The advances in biotechnological tools and the complete genome sequence of several citrus species will undoubtedly improve the breeding for citrus disease resistance with a much greater degree of precision. Here, we attempt to summarize the recent successful progress that has been achieved in the effective application of genetic engineering and genome editing technologies to obtain citrus disease-resistant (bacterial, fungal, and virus) crops. Furthermore, we also discuss the opportunities and challenges of genetic engineering and genome editing technologies for citrus disease resistance.
Citrus cultivars are propagated asexually through methods such as layering, cutting, and grafting. Grafting plays a critical role in regulating citrus fruit quality by optimizing stock–scion combinations. This study evaluated the grafting compatibility of two interstocks with Cocktail grapefruits (Citrus paradisi Macf. cv. Cocktail) by analyzing physiological and volatile traits of the fruits. We used Changshanhuyou (Citrus paradisi cv. Changshanhuyou) and Ponkan (Citrus reticulata Blanco cv. Ponkan), which are traditional citrus varieties in Quzhou, Zhejiang Province, as interstocks. Our results showed that the grapefruit fruits grafted onto Changshanhuyou exhibited higher total soluble solids (12.3 °Brix) and a brighter, more uniform peel color. The acidity of fruits grafted onto Changshanhuyou was lower than that of fruits grafted onto Ponkan and similar to the fruits from the control group that were grafted directly onto trifoliate orange. In terms of volatile organic compounds (VOCs), 974 and 577 unique VOCs were identified in the peel and pulp, respectively. In the peel, 34 (26 upregulated and 8 downregulated) and 14 (13 upregulated and 1 downregulated) differentially accumulated VOCs were identified when comparing the trifoliate orange group with Changshanhuyou and Ponkan groups, respectively. The pulp exhibited greater VOC diversity, with 272 (24 upregulated and 248 downregulated) and 199 (4 upregulated and 195 downregulated) differentially accumulated VOCs detected for the same comparisons. In conclusion, compared with direct grafting onto trifoliate orange or on Ponkan as an interstock, grafting onto Changshanhuyou resulted in better fruit quality and distinct differences in flesh flavor. These findings suggest that Changshanhuyou is a more suitable interstock for Cocktail grapefruit (Citrus paradisi Macf. cv. Cocktail) cultivation.