Key message: We found GmNAC06 plays an important role in salt stress responses through the phenotypic, physiological and molecular analyses of OE, VC, and Mutant composite soybean. Salinization affects 20% of all cultivated land worldwide because of the high salinity of irrigation water and the excessive use of water, and this amount is increasing daily. NAC (NAM, ATAF, and CUC) have been found to be involved in salt stress. In this study, a soybean NAC gene, GmNAC06 (Glyma06g21020.1), was cloned and functionally characterized. The results of expression analysis suggested that salt stress could influence the expression level of GmNAC06. The subcellular localization analysis results suggested that GmNAC06 may function as a transcription factor. Under salt stress, the overexpression technology combined with CRISPR-Cas9 system found that GmNAC06 could cause the accumulation of proline and glycine betaine to alleviate or avoid the negative effects of ROS; similarly, it could control the Na⁺/K⁺ ratios in hairy roots to maintain ionic homeostasis. The fresh weight of the transgenic hairy roots and the histochemical ROS staining of wild leaves suggested that transgenic hairy roots influence the function of wild leaves under salt stress conditions. Moreover, the expression levels of GmUBC2 and GmHKT1 were higher in the GmNAC06 hairy roots than in the control. Thus, the overexpression of GmNAC06 in hairy roots notably causes an entire composite plant to exhibit salt tolerance. The phenotype of composite soybean plants and transgenic Arabidopsis plants suggest that GmNAC06 plays a role in response to salt stress and could be useful in generating salt tolerant transgenic crops.

Kiwifruit (Actinidia chinensis Planch) is suitable for neutral acid soil. However, soil salinization is increasing in kiwifruit production areas, which has adverse effects on the growth and development of plants, leading to declining yields and quality. Therefore, analyzing the salt tolerance regulation mechanism can provide a theoretical basis for the industrial application and germplasm improvement of kiwifruit. We identified 120 NAC members and divided them into 13 subfamilies according to phylogenetic analysis. Subsequently, we conducted a comprehensive and systematic analysis based on the conserved motifs, key amino acid residues in the NAC domain, expression patterns, and protein interaction network predictions and screened the candidate gene AvNAC030. In order to study its function, we adopted the method of heterologous expression in Arabidopsis. Compared with the control, the overexpression plants had higher osmotic adjustment ability and improved antioxidant defense mechanism. These results suggest that AvNAC030 plays a positive role in the salt tolerance regulation mechanism in kiwifruit.

Grape (Vitis vinifera) is an important horticultural crop that can be used to make juice and wine. However, the small size of the berry limits its yield. Cultivating larger berry varieties can be an effective way to solve this problem. As the largest family of auxin early response genes, SAUR (small auxin upregulated RNA) plays an important role in the growth and development of plants. Berry size is one of the important factors that determine grape quality. However, the SAUR gene family’s function in berry size of grape has not been studied systematically. We identified 60 SAUR members in the grape genome and divided them into 12 subfamilies based on phylogenetic analysis. Subsequently, we conducted a comprehensive and systematic analysis on the SAUR gene family by analyzing distribution of key amino acid residues in the domain, structural features, conserved motifs, and protein interaction network, and combined with the heterologous expression in Arabidopsis and tomato. Finally, the member related to grape berry size in SAUR gene family were screened. This genome-wide study provides a systematic analysis of grape SAUR gene family, further understanding the potential functions of candidate genes, and provides a new idea for grape breeding.

Red coloration in apples, an important quality trait, is primarily attributed to the accumulation of anthocyanins. Centuries of breeding have produced a wide variety of apples with different levels of anthocyanins in response to genetic and environmental stimuli. The Huashuo apple shows a much darker red color than its sister line, Huarui. Thirteen different anthocyanins were detected in Huashuo and Huarui apples, of which ten were significantly more abundant in Huashuo apples, confirming that the color difference is indeed attributed to high anthocyanins accumulation rather than the types of anthocyanins. In particular, the contents of cyanidin 3-O-galactoside levels were highest among anthocyanins in both cultivars, reaching >5000 μg·g⁻¹ at the last color transition stage in Huashuo apples, while only >3000 μg·g⁻¹ in Huarui apples. Moreover, the expression of most structural genes, especially DFR, CHI, and 4CL associated with anthocyanin synthesis, were higher in Huashuo apples than in Huarui apples. Combined transcriptomics, metabolomics, and qRT-PCR analysis revealed that six transcription factors from the MYB and bZIP transcription factor families likely play key roles in the dark coloring of Huashuo apples. These results provide deeper insights into apple coloring and suggest a series of candidate genes for breeding anthocyanin-rich cultivars.

Kiwifruit is loved by consumers for its unique taste and rich vitamin C content. Kiwifruit are very sensitive to adverse soil environments owing to fleshy and shallow roots, which limits the uptake of water and nutrients into the root system, resulting in low yield and poor fruit quality. Lateral roots are the key organs for plants to absorb water and nutrients. Improving water and fertilizer use efficiency by promoting lateral root development is a feasible method to improve yield and quality. Expansin proteins plays a major role in lateral root growth; hence, it is important to identify expansin protein family members, screen key genes, and explore gene function in root development. In this study, 41 expansin genes were identified based on the genome of kiwifruit (‘Hongyang’, Actinidia chinensis). By clustering with the Arabidopsis thaliana expansin protein family, the 41 AcExpansin proteins were divided into four subfamilies. The AcExpansin protein family was further analysed by bioinformatics methods and was shown to be evolutionarily diverse and conserved at the DNA and protein levels. Based on previous transcriptome data and quantitative real-time PCR assays, we screened the candidate gene AcEXPA23. Overexpression of AcEXPA23 in kiwifruit increased the number of kiwifruit lateral roots.

The red color of apple peel is an important phenotypic and economic trait mainly attributed to anthocyanin accumulation. Apples show a deeper red color at higher altitudes than at lower ones; however, the molecular regulatory network underlying color variation along altitudinal gradients has not been investigated. In this study, the effects of environmental conditions associated with low (124 m) and high (1901 m) altitudes on peel color were assessed through physiological, metabolomic, transcriptomic, and qRT-PCR analyses in Huashuo apple and its sister line, Huarui apple. The content of cyanidin-3-O-galactoside, cyanidin-3-O-arabinoside, and cyanidin-3-O-xyloside was abundant in the high-altitude environment and may contribute to the deeper red color. Transcript levels of structural genes in the anthocyanin synthesis pathway, especially MdCHI, MdCHS, MdANS, and MdDFR, in apple peel were significantly higher at high altitude than at low altitude. Based on the protein interaction prediction and correlation analyses, four transcription factors (MDP0000127691, MDP0000284922, MDP0000758053, and MDP0000074681) could interact with anthocyanin synthesis-related proteins, showing high correlation with anthocyanin accumulation. Therefore, the abovementioned four genes and four transcription factors were predicted to account for the color differences between high and low altitudes. These results provide genetic resources and a theoretical basis for color-oriented fruit improvement.