The MYB transcription factor PpMYB5 regulates Pp4CL1/Pp4CL2 expression to promote lignin biosynthesis of fruit russeting in the flat nectarine.

KEY MESSAGE: Transcription factor PpMYB5 promotes lignin synthesis by directly binding to the Pp4CL1/Pp4CL2 promoter and affecting their expression, which may be related to nectarine russeting formation. Nectarine russeting is usually considered to be a non-invasive physiological disease that usually occurs on late-maturing cultivars and seriously affects their appearance quality and commercial value. The cause of nectarine fruit rust is currently unknown. In this study, we compared two flat nectarine cultivars, 'zhongyoupanweidi' (HD; russeting-free cultivar) and 'zhongyoupanweihou' (TH; russeting-prone cultivar), with respect to nectarine russeting by means of microscopy, transcriptomics, and hormone analysis. Compared to HD fruits, TH fruits had a broken cuticle, missing wax layer, and heavy lignin deposition. RNA sequencing (RNA-seq) revealed significant alternations in the expression of genes related to lignin synthesis. Moreover, structure genes Pp4CL1 and Pp4CL2, MYB transcription factor (TF) gene PpMYB5 were identified through weighted gene co-expression network analysis (WGCNA). Molecular experiments and transgenic evidence suggested that PpMYB5 regulates Pp4CL1/Pp4CL2 expression to promote lignin synthesis. Overall, in addition to providing new insights into the formation of mechanisms for nectarine russeting, our study also establishes a foundation for nectarine russeting prevention.

Machine learning-based identification of symptomatic carotid atherosclerotic plaques with dual-energy computed tomography angiography.

OBJECTIVE: This study aimed to develop and validate a machine learning model incorporating both dual-energy computed tomography (DECT) angiography quantitative parameters and clinically relevant risk factors for the identification of symptomatic carotid plaques to prevent acute cerebrovascular events. METHODS: The data of 180 patients with carotid atherosclerosis plaques were analysed from January 2017 to December 2021; 110 patients (64.03±9.58 years old, 20 women, 90 men) were allocated to the symptomatic group, and 70 patients (64.70±9.89 years old, 50 women, 20 men) were allocated to the asymptomatic group. Overall, five machine learning models using the XGBoost algorithm, based on different CT and clinical features, were developed in the training cohort. The performances of all five models were assessed in the testing cohort using receiver operating characteristic curves, accuracy, recall rate, and F1 score. RESULTS: The shapley additive explanation (SHAP) value ranking showed fat fraction (FF) as the highest among all CT and clinical features and normalised iodine density (NID) as the 10th. The model based on the top 10 features from the SHAP measurement showed optimal performance (area under the curve [AUC] .885, accuracy .833, recall rate .933, F1 score .861), compared with the other four models based on conventional CT features (AUC .588, accuracy .593, recall rate .767, F1 score .676), DECT features (AUC .685, accuracy .648, recall rate .667, F1 score .678), conventional CT and DECT features (AUC .819, accuracy .740, recall rate .867, F1 score .788), and all CT and clinical features (AUC .878, accuracy .833, recall rate .867, F1 score .852). CONCLUSION: FF and NID can serve as useful imaging markers of symptomatic carotid plaques. This tree-based machine learning model incorporating both DECT and clinical features could potentially comprise a non-invasive method for identification of symptomatic carotid plaques to guide clinical treatment strategies.

Grape Small Auxin Upregulated RNA (SAUR) 041 Is a Candidate Regulator of Berry Size in Grape.

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.

AvNAC030, a NAC Domain Transcription Factor, Enhances Salt Stress Tolerance in Kiwifruit.

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.

Type I H+-pyrophosphatase regulates the vacuolar storage of sucrose in citrus fruit.

The aim of this work was to evaluate the general role of the vacuolar pyrophosphatase proton pump (V-PPase) in sucrose accumulation in citrus species. First, three citrus V-PPase genes, designated CsVPP-1, CsVPP-2, and CsVPP-4, were identified in the citrus genome. CsVPP-1 and CsVPP-2 belonging to citrus type I V-PPase genes are targeted to the tonoplast, and CsVPP-4 belonging to citrus type II V-PPase genes is located in the Golgi bodies. Moreover, there was a significantly positive correlation between transcript levels of type I V-PPase genes and sucrose, rather than hexose, content in fruits of seven citrus cultivars. Drought and abscisic acid treatments significantly induced the CsVPP-1 and CsVPP-2 transcript levels, as well as the sucrose content. The overexpression of type I V-PPase genes significantly increased PPase activity, decreased pyrophosphate contents, and increased sucrose contents, whereas V-PPase inhibition produced the opposite effect in both citrus fruits and leaves. Furthermore, altering the expression levels of type I V-PPase genes significantly influenced the transcript levels of sucrose transporter genes. Taken together, this study demonstrated that CsVPP-1 and CsVPP-2 play key roles in sucrose storage in the vacuole by regulating pyrophosphate homeostasis, ultimately the sucrose biosynthesis and transcript levels of sucrose transport genes, providing a novel lead for engineering or breeding modified taste in citrus and other fruits.

Assessment of sugar and sugar accumulation-related gene expression profiles reveal new insight into the formation of low sugar accumulation trait in a sweet orange (Citrus sinensis) bud mutant.

The accumulation of soluble sugars in fleshy fruits largely determines their sweetness or taste. A spontaneous sweet orange mutant 'Hong Anliu' (HAL, Citrus sinensis) accumulates low soluble sugar content in fruit juice sacs than its wild type, 'Anliu' (AL) orange; however, the cause of reduced sugar content in 'HAL' fruit remains unclear. In this study, sugar content and expression profiles of genes involved in sugar metabolism and transport were compared between 'HAL' and 'AL' fruit juice sacs. In both cultivars, fructose and glucose displayed the increasing trends with significantly lower contents in 'HAL' than 'AL' after 160 DAF; moreover, sucrose had a declining trend in 'HAL' and increasing trend in 'AL' with fruit development. On the other hand, transcript levels of VINV, CWINV1, CWINV2, SUS4, SUS5, SPS1, SPS2, VPP-1, VPP-2, and some sugar transporter genes were significantly decreased in 'HAL' compared with 'AL' after 100 DAF or 160 DAF. Interestingly, the transcript levels of SPS2 and SUT2 exhibited a similar trend as it was found for sucrose content in both cultivars. These results suggested that the low sugar accumulation in 'HAL' fruit JS is accompanied by the reduced sink strength, sucrose-synthesis ability, and vacuolar storage ability compared with 'AL'; reduction of CWINVs, VINV, SPS2, SUT2, VPP-1, and VPP-2 transcript levels possibly plays a key role in the low storage of soluble sugars in the vacuoles of mutant juice sacs.

Positive-contrast susceptibility imaging based on first-order primal-dual optimization.

PURPOSE: To achieve faster reconstruction and better imaging quality of positive-contrast MRI based on the susceptibility mapping by incorporating a primal-dual (PD) formulation. METHODS: The susceptibility-based positive contrast MR technique was applied to estimate arbitrary magnetic susceptibility distributions of the metallic devices using a kernel deconvolution algorithm with a regularized ℓ1 minimization. The regularized positive-contrast inversion problem and its PD formulation were derived. The visualization of the positive contrast and convergence behavior of the PD algorithm were compared with those of the nonlinear conjugate gradient algorithm, fast iterative soft-thresholding algorithm, and alternating direction method of multipliers. These methods were tested and validated on computer simulations and phantom experiments. RESULTS: The PD approach could provide a faster reconstruction time compared with other methods. Experimental results showed that the PD algorithm could achieve comparable or even better visualization and accuracy of the metallic interventional devices in positive-contrast imaging with different SNRs and orientations to the B0 field. CONCLUSION: A susceptibility-based positive-contrast imaging technique by PD algorithm was proposed. The PD approach has more superior performance than other algorithms in terms of reconstruction time and accuracy for imaging the metallic interventional devices.

3D self-gated cardiac cine imaging at 3 Tesla using stack-of-stars bSSFP with tiny golden angles and compressed sensing.

PURPOSE: To develop and evaluate an accelerated 3D self-gated cardiac cine imaging technique at 3 Tesla without the use of external electrocardiogram triggering or respiratory gating. METHODS: A 3D stack-of-stars balanced steady-state free precession sequence with a tiny golden angle sampling scheme was developed to reduced eddy current effect-related artefacts at 3 Tesla. Respiratory and cardiac motion were derived from a central 5-point self-gating signal extraction approach. The data acquired around the end-expiration phases were then sorted into individual cardiac bins and used for reconstruction with compressed sensing. To evaluate the performance of the proposed method, image quality (1: the best; 4: the worst) was quantitatively compared using both the proposed method and the conventional 3D golden-angle self-gated method. Linear regression and Bland-Altman analysis were used to assess the functional measurements agreement between the proposed method and the routine 2D breath-hold multi-slice technique. RESULTS: Compared to the conventional 3D golden-angle self-gated method, the proposed method yielded images with much less streaking artifact and higher myocardium edge sharpness (0.50 ± 0.06 vs. 0.45 ± 0.05, P = 0.004). The proposed method provided an inferior image quality score to the routine 2D technique (2.13 ± 0.35 vs. 1.38 ± 0.52, P = 0.063) but a superior one to the conventional self-gated method (2.13 ± 0.35 vs. 3.13 ± 0.64, P = 0.031). Left ventricular functional measurements between the proposed method and routine 2D technique were all well in agreement. CONCLUSION: This study presents a novel self-gating approach to realize rapid 3D cardiac cine imaging at 3 Tesla.

Genome-wide identification of citrus ATP-citrate lyase genes and their transcript analysis in fruits reveals their possible role in citrate utilization.

ATP-citrate lyase (ACL, EC4.1.3.8) catalyzes citrate to oxaloacetate and acetyl-CoA in the cell cytosol, and has important roles in normal plant growth and in the biosynthesis of some secondary metabolites. We identified three ACL genes, CitACLα1, CitACLα2, and CitACLß1, in the citrus genome database. Both CitACLα1 and CitACLα2 encode putative ACL α subunits with 82.5 % amino acid identity, whereas CitACLß1 encodes a putative ACL ß subunit. Gene structure analysis showed that CitACLα1 and CitACLα2 had 12 exons and 11 introns, and CitACLß1 had 16 exons and 15 introns. CitACLα1 and CitACLß1 were predominantly expressed in flower, and CitACLα2 was predominantly expressed in stem and fibrous roots. As fruits ripen, the transcript levels of CitACLα1, CitACLß1, and/or CitACLα2 in cultivars 'Niuher' and 'Owari' increased, accompanied by significant decreases in citrate content, while their transcript levels decreased significantly in 'Egan No. 1' and 'Iyokan', although citrate content also decreased. In 'HB pummelo', in which acid content increased as fruit ripened, and in acid-free pummelo, transcript levels of CitACLα2, CitACLß1, and/or CitACLα1 increased. Moreover, mild drought stress and ABA treatment significantly increased citrate contents in fruits. Transcript levels of the three genes were significantly reduced by mild drought stress, and the transcript level of only CitACLß1 was significantly reduced by ABA treatment. Taken together, these data indicate that the effects of ACL on citrate use during fruit ripening depends on the cultivar, and the reduction in ACL gene expression may be attributed to citrate increases under mild drought stress or ABA treatment.

Identification and transcript analysis of two glutamate decarboxylase genes, CsGAD1 and CsGAD2, reveal the strong relationship between CsGAD1 and citrate utilization in citrus fruit.

Glutamate decarboxylase (GAD, EC 4.1.1.15) has been suggested to be a key, regulatory point in the biosynthesis of γ-aminobutyrate (GABA) and in the utilization of citric acid through GABA shunt pathway. In this study we discovered two GAD genes, named as CsGAD1 and CsGAD2, in citrus genome database and then successfully cloned. Both CsGAD1 and CsGAD2 have a putative pyridoxal 5-phosphate binding domain in the middle region and a putative calmodulin-binding domain at the carboxyl terminus. Gene structure analysis showed that much difference exists in the size of exons and introns or in cis-regulatory elements in promoter region between the two GAD genes. Gene expression indicated that CsGAD1 transcript was predominantly expressed in flower and CsGAD2 transcript was predominantly expressed in fruit juice sacs; in the ripening fruit, CsGAD1 transcript level was at least 2-time higher than CsGAD2 transcript level. Moreover, CsGAD1 transcript level was increased significantly along with the increase of GAD activity and accompanied by a significant decrease of titratable acid (TA), suggesting that it is CsGAD1 rather than CsGAD2 plays a role in the citric acid utilization during fruit ripening. In addition, injection of abscisic acid and foliar spray of K2SO4 significantly increased the TA content of Satsuma mandarin, and significantly decreased GAD activity as well as CsGAD1 transcript, further suggesting the important role of CsGAD1 in the citrate utilization of citrus fruit.