Summary: Ethylene (ETH) controls climacteric fruit ripening and can be triggered by osmotic stress. However, the mechanism regulating ETH biosynthesis during fruit ripening and under osmotic stress is largely unknown in apple (Malus domestica). Here, we explored the roles of SnRK2 protein kinases in ETH biosynthesis related to fruit ripening and osmoregulation. We identified the substrates of MdSnRK2‐I using phosphorylation analysis techniques. Finally, we identified the MdSnRK2‐I‐mediated signaling pathway for ETH biosynthesis related to fruit ripening and osmoregulation. The activity of two MdSnRK2‐I members, MdSnRK2.4 and MdSnRK2.9, was significantly upregulated during ripening or following mannitol treatment. Overexpression of MdSnRK2‐I increased ETH biosynthesis under normal and osmotic conditions in apple fruit. MdSnRK2‐I phosphorylated the transcription factors MdHB1 and MdHB2 to enhance their protein stability and transcriptional activity on MdACO1. MdSnRK2‐I also interacted with MdACS1 and increased its protein stability through two phosphorylation sites. The increased MdACO1 expression and MdACS1 protein stability resulted in higher ETH production in apple fruit. In addition, heterologous expression of MdSnRK2‐I or manipulation of SlSnRK2‐I expression in tomato (Solanum lycopersicum) fruit altered fruit ripening and ETH biosynthesis. We established that MdSnRK2‐I functions in fruit ripening and osmoregulation, and identified the MdSnRK2‐I‐mediated signaling pathway controlling ETH biosynthesis.

Strawberry is increasingly used as a model plant for research on fruit growth and development. The transient gene manipulation (TGM) technique is widely used to determine the function of plant genes, including those in strawberry fruits. However, its reliable application for the precise identification of gene function has been difficult owing to the lack of conditional optimization. In this study, we found that successful transient gene manipulation requires optimization, with the vector type, temperature, and fruit developmental stage being three major factors determining success. Notably, we found that transient gene manipulation was feasible only from the large green fruit stage onwards, making it especially suitable for identifying genes involved in strawberry fruit ripening. Furthermore, we established a method called percentage difference of phenotype (PDP), in which the functional effect of a gene could be precisely and efficiently identified in strawberry fruits. This method can be used to estimate the functional effect of a gene as a value from 0 to 100%, such that different genes can be quantitatively compared for their relative abilities to regulate fruit ripening. This study provides a useful tool for accelerating research on the molecular basis of strawberry fruit ripening.

Abscisic acid (ABA) plays a major role in the regulation of strawberry fruit ripening; however, the origin of the ABA signal is largely unknown. Here, we report an autocatalytic mechanism for ABA biosynthesis and its synergistic interaction with auxin to regulate strawberry fruit ripening. We demonstrate that ABA biosynthesis is self-induced in the achenes but not in the receptacle, resulting in its substantial accumulation during ripening. ABA was found to regulate both IAA transport and biosynthesis, thereby modulating IAA content during both early fruit growth and later fruit ripening. Taken together, these results reveal the origins of the ABA signal and demonstrate the importance of its coordinated action with IAA in the regulation of strawberry fruit development and ripening.

Plant cell walls are essential structures for plant growth and development as well as plant adaptation to environmental stresses. Thus, plants have evolved signaling mechanisms to monitor the changes in the cell wall structure, triggering compensatory changes to sustain cell wall integrity (CWI). CWI signaling can be initiated in response to environmental and developmental signals. However, while environmental stress-associated CWI signaling has been extensively studied and reviewed, less attention has been paid to CWI signaling in relation to plant growth and development under normal conditions. Fleshy fruit development and ripening is a unique process in which dramatic alternations occur in cell wall architecture. Emerging evidence suggests that CWI signaling plays a pivotal role in fruit ripening. In this review, we summarize and discuss the CWI signaling in relation to fruit ripening, which will include cell wall fragment signaling, calcium signaling, and NO signaling, as well as Receptor-Like Protein Kinase (RLKs) signaling with an emphasis on the signaling of FERONIA and THESEUS, two members of RLKs that may act as potential CWI sensors in the modulation of hormonal signal origination and transduction in fruit development and ripening.

Fleshy fruit ripening involves changes in numerous cellular processes and metabolic pathways, resulting from the coordinated actions of diverse classes of structural and regulatory proteins. These include enzymes, transporters and complex signal transduction systems. Many aspects of the signaling machinery that orchestrates the ripening of climacteric fruits, such as tomato (Solanum lycopersicum), have been elucidated, but less is known about analogous processes in non-climacteric fruits. The latter include strawberry (Fragaria x ananassa) and grape (Vitis vinifera), both of which are used as non-climacteric fruit experimental model systems, although they originate from different organs: the grape berry is a true fruit derived from the ovary, while strawberry is an accessory fruit that is derived from the floral receptacle. In this article, we summarize insights into the signal transduction events involved in strawberry and grape berry ripening. We highlight the mechanisms underlying non-climacteric fruit ripening, the multiple primary signals and their integrated action, individual signaling components, pathways and their crosstalk, as well as the associated transcription factors and their signaling output.

Fleshy fruits have been traditionally categorized into climacteric (CL) and non-climacteric (NC) groups. CL fruits share a common ripening mechanism of hormonal regulation, i.e., the ethylene regulation, whereas whether NC fruits share a common mechanism remains controversial. Abscisic acid (ABA) has been commonly thought to be a key regulator in NC fruit ripening; however, besides ABA, many other hormones have been increasingly suggested to play crucial roles in NC fruit ripening. NC fruits vary greatly in their organ origin, constitution, and structure. Development of different organs may be different in the pattern of hormonal regulation. It has been well demonstrated that the growth and development of strawberry, the model of NC fruits, is largely controlled by a hormonal communication between the achenes and receptacle; however, not all NC fruits contain achenes. Accordingly, it is particularly important to understand whether strawberry is indeed able to represent a universal mechanism for the hormonal regulation of NC fruit ripening. In this mini-review, we summarized the recent research advance on the hormone regulation of NC ripening in relation to fruit organ origination, constitution, and structure, whereby analyzing and discussing whether NC fruits may share a common mechanism of hormonal regulation.