Integrated metabolic and transcriptional analysis reveals the role of carotenoid cleavage dioxygenase 4 (IbCCD4) in carotenoid accumulation in sweetpotato tuberous roots.

BACKGROUND: Plant carotenoids are essential for human health, having wide uses in dietary supplements, food colorants, animal feed additives, and cosmetics. With the increasing demand for natural carotenoids, plant carotenoids have gained great interest in both academic and industry research worldwide. Orange-fleshed sweetpotato (Ipomoea batatas) enriched with carotenoids is an ideal feedstock for producing natural carotenoids. However, limited information is available regarding the molecular mechanism responsible for carotenoid metabolism in sweetpotato tuberous roots. RESULTS: In this study, metabolic profiling of carotenoids and gene expression analysis were conducted at six tuberous root developmental stages of three sweetpotato varieties with different flesh colors. The correlations between the expression of carotenoid metabolic genes and carotenoid levels suggested that the carotenoid cleavage dioxygenase 4 (IbCCD4) and 9-cis-epoxycarotenoid cleavage dioxygenases 3 (IbNCED3) play important roles in the regulation of carotenoid contents in sweetpotato. Transgenic experiments confirmed that the total carotenoid content decreased in the tuberous roots of IbCCD4-overexpressing sweetpotato. In addition, IbCCD4 may be regulated by two stress-related transcription factors, IbWRKY20 and IbCBF2, implying that the carotenoid accumulation in sweeetpotato is possibly fine-tuned in responses to stress signals. CONCLUSIONS: A set of key genes were revealed to be responsible for carotenoid accumulation in sweetpotato, with IbCCD4 acts as a crucial player. Our findings provided new insights into carotenoid metabolism in sweetpotato tuberous roots and insinuated IbCCD4 to be a target gene in the development of new sweetpotato varieties with high carotenoid production.

Retraction notice to "Genome-wide analysis of the B-box gene family in the sweet potato wild ancestor Ipomoea trifida and determination of the function of IbBBX28 in the regulation of flowering time of Arabidopsis" [Plant Physiol. Biochem. 188 (2022) 109-122].

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editors-in-Chief. A large part of the article is highly similar to the paper previously published by Wenqian Hou, Lei Ren, Yang Zhang, Haoyun Sun, Tianye Shi, Yulan Gu, Aimin Wang, Daifu Ma, Zongyun Li and Lei Zhang in Scientia Horticulturae 288 (2021) 110374 https://doi.org/10.1016/j.scienta.2021.110374. In particular, a large part of the two articles shows a study on the same gene family in the same plant, with similar methodological approaches, resulting in a series of highly similar figures. One of the conditions of submission of a paper for publication is that authors declare explicitly that their work is original and has not appeared in a publication elsewhere. Re-use of any data should be appropriately cited. As such this article represents a severe abuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of the journal that this was not detected during the submission process.

The sweet potato B-box transcription factor gene IbBBX28 negatively regulates drought tolerance in transgenic Arabidopsis.

B-box (BBX) which are a class of zinc finger transcription factors, play an important role in regulating of photoperiod, photomorphogenesis, and biotic and abiotic stresses in plants. However, there are few studies on the involvement of BBX transcription factors in response to abiotic stresses in sweet potato. In this paper, we cloned the DNA and promoter sequences of IbBBX28. There was one B-box conserved domain in IbBBX28, and the expression of IbBBX28 was induced under drought stress. Under drought stress, compared to wild type Arabidopsis, the protective enzyme activities (SOD, POD, and CAT) were all decreased in IbBBX28-overexpression Arabidopsis but increased in the mutant line bbx28, while the MDA content was increased in the IbBBX28-overexpression Arabidopsis and decreased in the bbx28. Moreover, the expression levels of the resistance-related genes showed the same trend as the protective enzyme activities. These results showed that IbBBX28 negatively regulates drought tolerance in transgenic Arabidopsis. Additionally, the yeast two-hybrid and BiFC assays verified that IbBBX28 interacted with IbHOX11 and IbZMAT2. The above results provide important clues for further studies on the role of IbBBX28 in regulating the stress response in sweet potato.

Overexpression of Sweet Potato Carotenoid Cleavage Dioxygenase 4 (IbCCD4) Decreased Salt Tolerance in Arabidopsis thaliana.

Salt stress has a serious impact on normal plant growth and yield. Carotenoid cleavage dioxygenase (CCD) degrades carotenoids to produce apocarotenoids, which are involved in plant responses to biotic and abiotic stresses. This study shows that the expression of sweet potato IbCCD4 was significantly induced by salt and dehydration stress. The heterologous expression of IbCCD4 in Arabidopsis was induced to confirm its salt tolerance. Under 200 mM NaCl treatment, compared to wild-type plants, the rosette leaves of IbCCD4-overexpressing Arabidopsis showed increased anthocyanins and carotenoid contents, an increased expression of most genes in the carotenoid metabolic pathway, and increased malondialdehyde (MDA) levels. IbCCD4-overexpressing lines also showed a decreased expression of resistance-related genes and a lower activity of three antioxidant enzymes: peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT). These results indicate that IbCCD4 reduced salt tolerance in Arabidopsis, which contributes to the understanding of the role of IbCCD4 in salt stress.

Genome-wide analysis of the B-box gene family in the sweetpotato wild ancestor Ipomoea trifida and determination of the function of IbBBX28 in the regulation of flowering time of Arabidopsis.

B-box (BBX) proteins constitute a class of transcription factors that play vital roles in the regulation of photoperiod flowering, photomorphogenesis, and the response to biotic and abiotic stresses. In this paper, a total of 32 BBX genes were identified in Ipomoea trifida, a wild ancestor of sweetpotato. Chromosome localization analysis showed that these 32 ItfBBX genes were distributed unevenly across 12 chromosomes. The ItfBBX gene family members were classified into five groups according to their phylogenetic relationships and structural features. Predictions of cis-elements revealed that the promoter sequences of the ItfBBX genes contain light response, stress response, hormone response and other elements. Synteny analysis revealed evidence of 26 segmental duplication events and only one tandem duplication event. Tissue-specific and abiotic stress-response expression profiles were analysed, and the results were confirmed via RT-qPCR. Overall, ItfBBX genes may play vital roles in the stress response. We chose IbBBX28 for further study and revealed that IbBBX28 negatively regulates the flowering time of IbBBX28-overexpressing Arabidopsis under long-day conditions. Our study provides references for characterizing the function of BBX genes in sweetpotato.