Genome-wide identification of the AcMADS-box family and functional validation of AcMADS32 involved in carotenoid biosynthesis in Actinidia.

MADS-box is a large transcription factor family in plants and plays a crucial role in various plant developmental processes; however, it has not been systematically analyzed in kiwifruit. In the present study, 74 AcMADS genes were identified in the Red5 kiwifruit genome, including 17 type-I and 57 type-II members according to the conserved domains. The AcMADS genes were randomly distributed across 25 chromosomes and were predicted to be mostly located in the nucleus. A total of 33 fragmental duplications were detected in the AcMADS genes, which might be the main force driving the family expansion. Many hormone-associated cis-acting elements were detected in the promoter region. Expression profile analysis showed that AcMADS members had tissue specificity and different responses to dark, low temperature, drought, and salt stress. Two genes in the AG group, AcMADS32 and AcMADS48, had high expression levels during fruit development, and the role of AcMADS32 was further verified by stable overexpression in kiwifruit seedlings. The content of α-carotene and the ratio of zeaxanthin/ß-carotene was increased in transgenic kiwifruit seedlings, and the expression level of AcBCH1/2 was significantly increased, suggesting that AcMADS32 plays an important role in regulating carotenoid accumulation. These results have enriched our understanding of the MADS-box gene family and laid a foundation for further research of the functions of its members during kiwifruit development.

AcMADS32 positively regulates carotenoid biosynthesis in kiwifruit by activating AcBCH1/2 expression.

Fruits provide abundant carotenoid nutrients for humans, whereas the understanding of the transcriptional regulatory mechanisms of carotenoids in fruits is still limited. Here, we identified a transcription factor AcMADS32 in kiwifruit, which was highly expressed in the fruit, correlated with carotenoid content and localized in the nucleus. The silencing expression of AcMADS32 significantly reduced the content of ß-carotene and zeaxanthin and expression of ß-carotene hydroxylase gene AcBCH1/2 in kiwifruit, while transient overexpression increased the accumulation of zeaxanthin, suggesting that AcMADS32 was an activator involved in the transcriptional regulation of carotenoid in fruit. When AcMADS32 was further stably transformed into kiwifruit, the content of total carotenoid and components in the leaves of transgenic lines significantly increased, and the expression level of carotenogenic genes was up-regulated. Moreover, Y1H and dual luciferase reporter experiments confirmed that AcMADS32 directly bound the AcBCH1/2 promoter and activated its expression. Through Y2H assays, AcMADS32 can interact with other MADS transcription factor AcMADS30, AcMADS64 and AcMADS70. These findings will contribute to our understanding of the transcriptional regulation mechanisms underlying carotenoid biosynthesis in plants.

Significant increases in Donghong kiwifruit yield by a novel umbrella-shaped trellis system and identification of associated molecular mechanisms.

China is the largest kiwifruit producer in the world, accounting for more than half of the total. However, in terms of yield per unit area, China is much lower than the global average and lags behind that of other countries. Yield improvement is of critical importance for the current kiwifruit industry in China. In this study, an improved overhead pergolas trellis (OPT) system, namely, the umbrella-shaped trellis (UST) system, was developed for Donghong kiwifruit, which is now the second most popular and widely cultivated red-fleshed kiwifruit in China. Surprisingly, the estimated yield on the UST system was more than two times higher than that with a traditional OPT, while the external fruit quality was maintained and the internal fruit quality was improved. One of the mechanisms contributing to the yield improvement was the significant promotion of the vegetative growth of canes at 6 ~ 10 mm in diameter by the UST system. The upper canopy of the UST treatment served as a natural shading condition for the lower fruiting canopy and thus had positive effects on the accumulation of chlorophylls and total carotenoids in the fruiting canopy. The most productive zones on the fruiting canes (6 ~ 10 mm in diameter) contained significantly higher (P < 0.05) levels of zeatin riboside (ZR) and auxin (IAA) and ratios of ZR/gibberellin (GA), ZR/abscisic acid (ABA), and ABA/GA. A relatively high carbon/nitrogen ratio may promote the flower bud differentiation process of Donghong kiwifruit. The outcomes of this study provide a scientific basis for manifold increase in production of kiwifruit and contribute to the sustainability of the kiwifruit industry.

Melatonin and arbuscular mycorrhizal fungi synergistically improve drought toleration in kiwifruit seedlings by increasing mycorrhizal colonization and nutrient uptake.

Kiwifruit is a vine fruit tree that is vulnerable to water deficiency due to its shallow root system and large leaves. Although mycorrhizal inoculation and melatonin application has been proved to improve plants drought tolerance, their interaction effects are still unclear. In this study, arbuscular mycorrhizal (AM) fungi incubation and melatonin (MT) irrigation were applied to kiwifruit seedlings alone or in combination to investigate their effect on drought tolerance. The results revealed that AM had more effect on promoting root biomass, water use efficiency, and uptake of nitrogen, phosphorus and iron. While MT was more effective in promoting shoot biomass and antioxidant enzyme activities to remove reactive oxygen species accumulation. Moreover, MT supplementary significantly increased the AM colonization, spore density and hyphal length density in roots. Therefore, combined application of AM fungi and MT had additive effects on improvement biomass accumulation, increasing chlorophyll content, photosynthetic efficiency, catalase activity, and decreasing malondialdehyde accumulation under drought stress, thus promoting plant growth and alleviating the drought damage to plant. These results provide guidance for AM and MT combined application to improve abiotic resistance in plants.