Genome­wide analysis of the GT8 gene family in apple and functional identification of MhGolS2 in saline-alkali tolerance.

Members of the glycosyltransferase 8 (GT8) family play an important role in regulating gene expression in response to many kinds of biotic and abiotic stress. In this study, 56 members of the apple GT8 family were identified, and their gene structure, phylogenetic relationships, chromosomal localization, and promoter cis-acting elements were comprehensively analyzed. Subsequently, 20 genes were randomly selected from the evolutionary tree for qRT-PCR detection, and it was found that MhGolS2 was significantly overexpressed under stress conditions. MhGolS2 was isolated from M.halliana and transgenic Arabidopsis thaliana, tobacco and apple callus tissues were successfully obtained. The transgenic plants grew better under stress conditions with higher polysaccharide, chlorophyll and proline content, lower conductivity and MDA content, significant increase in antioxidant enzyme activities (SOD, POD, CAT) and maintenance of low Na+/K+ as compared to the wild type. Meanwhile, the expression levels of reactive oxygen species-related genes (AtSOD, AtPOD, and AtCAT), Na+ transporter genes (AtCAX5, AtSOS1, and AtHKT1), H+-ATPase genes (AtAHA2 and AtAHA8), and raffinose synthesis-related genes (AtSTS, AtRFS1, and AtMIPS) were significantly up-regulated, while the expression levels of K+ transporter genes (AtSKOR, AtHAK5) were reduced. Finally, the Y2H experiment confirmed the interaction between MhGolS2 and MhbZIP23, MhMYB1R1, MhbHLH60, and MhNAC1 proteins. The above results indicate that MhGolS2 can improve plant saline-alkali tolerance by promoting polysaccharide synthesis, scavenging reactive oxygen species, and increasing the activity of antioxidant enzymes. This provides excellent stress resistance genes for the stress response regulatory network in apple.

Transcription factors and plant hormones mediate wax metabolism in response to drought stress.

Plants have, throughout evolution, developed a hydrophobic cuticle to protect them from various stresses in the terrestrial environment. The cuticle layer is mainly composed of cutin and cuticular wax, a mixture of very-long-chain fatty acids and their derivatives. With the progress of transcriptome sequencing and other research methods, the key enzymes, transporters and regulatory factors in wax synthesis and metabolism have been gradually identified, especially the study on the regulation of wax metabolism by transcription factors and others in response to plant stress has become a hot topic. Drought is a major abiotic stress that limits plant growth and crop productivity. Plant epidermal wax prevents non-stomatal water loss and improves water use efficiency to adapt to arid environments. In this study, the ways of wax synthesis, transport, metabolism and regulation at different levels are reviewed. At the same time, the regulation of wax by different transcription factors and plant hormones in response to drought is elaborated, and key research questions and important directions for future solutions are proposed to enhance the potential application of epidermal wax in agriculture and the environment.

Genome-wide identification and expression analysis of the ALDH gene family and functional analysis of PaALDH17 in Prunus avium.

ALDH (Aldehyde dehydrogenase), as an enzyme that encodes the dehydroxidization of aldehydes into corresponding carboxylic acids, played an important role inregulating gene expression in response to many kinds of biotic and abiotic stress, including saline-alkali stress. Saline-alkali stress was a common stress that seriously affected plant growth and productivity. Saline-alkali soil contained the characteristics of high salinity and high pH value, which could cause comprehensive damage such as osmotic stress, ion toxicity, high pH, and HCO3-/CO32- stress. In our study, 18 PaALDH genes were identified in sweet cherry genome, and their gene structures, phylogenetic analysis, chromosome localization, and promoter cis-acting elements were analyzed. Quantitative real-time PCR confirmed that PaALDH17 exhibited the highest expression compared to other members under saline-alkali stress. Subsequently, it was isolated from Prunus avium, and transgenic A. thaliana was successfully obtained. Compared with wild type, transgenic PaALDH17 plants grew better under saline-alkali stress and showed higher chlorophyll content, Superoxide dismutase (SOD), Peroxidase (POD) and Catalase (CAT) enzyme activities, which indicated that they had strong resistance to stress. These results indicated that PaALDH17 improved the resistance of sweet cherries to saline-alkali stress, which in turn improved quality and yields. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01444-7.

Molecular cloning and functional characterization in response to saline-alkali stress of the MhZEP gene in Arabidopsis thaliana.

Soil salinization is one of the major environmental factors that restrict plant growth and development. Zeaxanthin epoxidase (ZEP) functions in ABA biosynthesis and the xanthophyll cycle and has a vital role in plant responses to various environmental stresses. It was found by quantitative real-time PCR (qRT-PCR) that MhZEP responded to saline-alkali stress and showed the highest expression at 48 h of saline-alkali stress, which was 14.53-fold of 0 h. The MhZEP gene was cloned from the apple rootstock begonia (Malus halliana Koehne) and its protein physicochemical properties were analyzed. Subsequently, the functional characterization of MhZEP (ID: 103403091) was further investigated in Arabidopsis thaliana. The MhZEP contained a complete open reading frame with a length of 1998 bp, and encoded 665 amino acids with an isoelectric point of 7.18. Phylogenetic tree analysis showed that MhZEP was the most homologous and closely related to Glycine max. Compared with wild-type, transgenic plants grew better under saline-alkali stress and the MhZEP-OE line showed higher chlorophyll content, carotenoid content, enzyme activities (POD, SOD, CAT and APX) and K+ content, whereas they had lower chlorosis and Na+ content than the wild type (WT), which indicated that they had strong resistance to stress. The expression levels of saline-alkali stress-related genes in A. thaliana MhZEP-OE were examined by qRT-PCR, and it was found that the MhZEP improved the tolerance of A. thaliana to saline-alkali stress tolerance by regulating the expression of carotenoid synthesis genes (MhPSY, MhZDS, MhLYCB and MhVDE) and ABA biosynthesis genes (MhNCED5, MhABI1 and MhCYP707A2). And the potassium-sodium ratio in the cytoplasm was increased to maintain ionic homeostasis by modulating the expression of Na+ transporter genes (MhCHX15 and MhSOS1) and K+ transporter genes (MhHKT1;1, MhNHX1 and MhSKOR1). Moreover, the expression of H+-ATPase genes (MhAHA2 and MhAHA8) was increased to reduce the oxidative damage caused by saline-alkali stress. In summary, MhZEP acted as an essential role in plant resistance to saline-alkali stress, which lays the foundation for further studies on its function in apple.

Transcriptomic analysis reveals GA3 is involved in regulating flavonoid metabolism in grape development for facility cultivation.

Gibberellin, as one of the pivotal plant growth regulators, can improve fruit quality by altering fruit size and secondary metabolite content. Flavonoids are the most abundant secondary metabolites in grapes, which influence the color and quality of the fruit. However, the molecular mechanism of whether and how GA3 affects flavonoid metabolism has not been reported, especially for the 'Red globe' grape with delayed cultivation in Hexi corridor. In the present study, the 'Red globe' grape grown in delayed facilities was sprayed with 20, 40, 60, 80 and 100 mg/L GA3 at berries pea size (BPS), veraison (V) and berries ripe (BR), respectively. The results showed that the berry weight, soluble sugar content and secondary metabolite content (the flavonoid content, anthocyanin content and polyphenol content) at BR under 80 mg/L GA3 treatment were remarkably increased compared with other concentration treatments. Therefore, RNA sequencing (RNA-seq) was used to analyze the differentially expressed genes (DEGS) and pathways under 80 mg/L GA3 treatment at three periods. GO analysis showed that DEGs were closely related to transporter activity, cofactor binding, photosynthetic membrane, thylakoid, ribosome biogenesis and other items. The KEGG enrichment analysis found that the DEGs were mainly involved in flavonoid biosynthesis and phenylpropanoid biosynthesis, indicating GA3 exerted an impact on the color and quality of berries through these pathways. In conclusion, GA3 significantly increased the expression of genes related to flavonoid synthesis, enhanced the production of secondary metabolites, and improved fruit quality. In addition, these findings can provide a theoretical basis for GA3 to modulate the accumulation and metabolism of flavonoids in grape fruit.

Transcription factors dealing with Iron-deficiency stress in plants: focus on the bHLH transcription factor family.

Iron (Fe), as an important micronutrient element necessary for plant growth and development, not only participates in multiple physiological and biochemical reactions in cells but also exerts a crucial role in respiration and photosynthetic electron transport. Since Fe is mainly present in the soil in the form of iron hydroxide, Fe deficiency exists universally in plants and has become an important factor triggering crop yield reduction and quality decline. It has been shown that transcription factors (TFs), as an important part of plant signaling pathways, not only coordinate the internal signals of different interaction partners during plant development, but also participate in plant responses to biological and abiotic stresses, such as Fe deficiency stress. Here, the role of bHLH transcription factors in the regulation of Fe homeostasis (mainly Fe uptake) is discussed with emphasis on the functions of MYB, WRKY and other TFs in the maintenance of Fe homeostasis. This review provides a theoretical basis for further studies on the regulation of TFs in Fe deficiency stress response.

Functional identification of ZDS gene in apple (Malus halliana) and demonstration of it's role in improving saline-alkali stress tolerance.

Carotenoids are powerful antioxidants that mediate transfer of electrons, directly affect abiotic stress responses in plants through regulating activity of antioxidant enzymes. ζ-Carotene desaturase (ZDS) is a key enzyme in carotenoid biosynthesis pathway, which can catalyze ζ-carotene to form lycopene to regulate carotenoid biosynthesis and accumulation. However, the mechanism of its regulation of saline-alkali stress remains unclear. In this research, based on transcriptomic analysis of Malus halliana with a apple rootstock, we screened out ZDS gene (LOC103451012), with significantly high expression by saline-alkali stress, whose expression in the leaves was 10.8-fold than that of the control (0 h) under 48 h of stress. Subsequently, the MhZDS gene was isolated from M. halliana, and transgenic Arabidopsis thaliana, tobacco, and apple calli were successfully obtained through agrobacterium-mediated genetic transformation. We found that overexpression of MhZDS enhanced the tolerance of A. thaliana, tobacco and apple calli under saline-alkali stress and caused a variety of physiological and biochemical changes: compared with wild-type, transgenic plants grew better under saline stress and MhZDS-OE lines showed higher chlorophyll content, POD, SOD, CAT activities and proline content, lower electrical conductivity and MDA content. These results indicate that MhZDS plays an important role in plant resistance to saline-alkali stress, providing excellent resistance genes for the regulatory network of salinity stress response in apples and provide a theoretical basis for the breeding of apple varieties with strong saline-alkali resistance. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01333-5.

Associations of combined healthy lifestyles with cancer morbidity and mortality among individuals with diabetes: results from five cohort studies in the USA, the UK and China.

AIMS/HYPOTHESIS: Cancer has contributed to an increasing proportion of diabetes-related deaths, while lifestyle management is the cornerstone of both diabetes care and cancer prevention. We aimed to evaluate the associations of combined healthy lifestyles with total and site-specific cancer risks among individuals with diabetes. METHODS: We included 92,239 individuals with diabetes but without cancer at baseline from five population-based cohorts in the USA (National Health and Nutrition Examination Survey and National Institutes of Health [NIH]-AARP Diet and Health Study), the UK (UK Biobank study) and China (Dongfeng-Tongji cohort and Kailuan study). Healthy lifestyle scores (range 0-5) were constructed based on current nonsmoking, low-to-moderate alcohol drinking, adequate physical activity, healthy diet and optimal bodyweight. Cox regressions were used to calculate HRs for cancer morbidity and mortality, adjusting for sociodemographic, medical and diabetes-related factors. RESULTS: During 376,354 person-years of follow-up from UK Biobank and the two Chinese cohorts, 3229 incident cancer cases were documented, and 6682 cancer deaths were documented during 1,089,987 person-years of follow-up in the five cohorts. The pooled multivariable-adjusted HRs (95% CIs) comparing participants with 4-5 vs 0-1 healthy lifestyle factors were 0.73 (0.61, 0.88) for incident cancer and 0.55 (0.46, 0.67) for cancer mortality, and ranged between 0.41 and 0.63 for oesophagus, lung, liver, colorectum, breast and kidney cancers. Findings remained consistent across different cohorts and subgroups. CONCLUSIONS/INTERPRETATION: This international cohort study found that adherence to combined healthy lifestyles was associated with lower risks of total cancer morbidity and mortality as well as several subtypes (oesophagus, lung, liver, colorectum, breast and kidney cancers) among individuals with diabetes.

Genome-wide identification of the CER1 gene family in apple and response of MdCER1-1 to drought stress.

Plant cuticular wax was a major consideration affecting the growth and quality of plants through protecting the plant from drought and other diseases. According to existing studies, CER1, as the core enzyme encoding the synthesis of alkanes, the main component of wax, can directly affect the response of plants to stress. However, there were few studies on the related functions of CER1 in apple. In this study, three MdCER1 genes in Malus domestica were identified and named MdCER1-1, MdCER1-2, and MdCER1-3 according to their distribution on chromosomes. Then, their physicochemical properties, sequence characteristics, and expression patterns were analyzed. MdCER1-1, with the highest expression level among the three members, was screened for cloning and functional verification. Real-time fluorescence quantitative PCR (qRT-PCR) analysis also showed that drought stress could increase the expression level of MdCER1-1. The experiment of water loss showed that overexpression of MdCER1-1 could effectively prevent water loss in apple calli, and the effect was more significant under drought stress. Meanwhile, MdYPB5, MdCER3, and MdKCS1 were significantly up-regulated, which would be bound up with waxy metabolism. Gas chromatography-mass spectrometer assay of wax fraction makes known that overexpression of MdCER1-1 apparently scaled up capacity of alkanes. The enzyme activities (SOD, POD) of overexpressed apple calli increased significantly, while the contents of proline increased compared with wild-type calli. In conclusion, MdCER1-1 can resist drought stress by reducing water loss in apple calli epidermis, increasing alkanes component content, stimulating the expression of waxy related genes (MdYPB5, MdCER3, and MdKCS1), and increasing antioxidant enzyme activity, which also provided a theoretical basis for exploring the role of waxy in other stresses.

Saline-alkali stress tolerance is enhanced by MhPR1 in Malus halliana leaves as shown by transcriptomic analyses.

MAIN CONCLUSION: qRT-PCR analysis showed that MhPR1 was strongly induced by saline-alkali stress. Overexpression of MhPR1 enhanced tolerance to saline-alkali stress in transgenic tobacco (Nicotiana tabacum L.) and apple calli. Abstract: Soil salinization seriously threaten apple growth in Northwest loess plateau of China. Malus halliana has developed special system to adapt to saline-alkali environmental stress. To obtain a more detailed understanding of the adaptation mechanisms involved in M. halliana, a transcriptomic approach was used to analyze the leaves' pathways in the stress and its regulatory mechanisms. RNA-Seq showed that among the 16,246 investigated unigenes under saline-alkali stress, 7268 genes were up-regulated and 8978 genes were down-regulated. KEGG analysis indicated that most of the enriched saline-alkali-responsive genes were mainly involved in plant hormone, calcium signal transduction, amino acids, carotenoid and flavonoids biosynthesis, carbon and phenylalanine metabolism, and other secondary metabolites. Expression profile analysis by quantitative real-time PCR confirmed that the maximum up-regulation of MhPR1 under saline-alkali stress was 7.1 folds in leaves. Overexpression of MhPR1 enhanced tolerance to saline-alkali stress in transgenic tobacco (Nicotiana tabacum L.) and apple calli. Taken together, our results demonstrate that MhPR1 encodes a saline-alkali-responsive transcriptional activator and provide valuable information for further study of PR1 functions in apple.

Dietary Approaches to Stop Hypertension (DASH)-Style Dietary Pattern and 24-Hour Ambulatory Blood Pressure in Elderly Chinese with or without Hypertension.

BACKGROUND: It is unknown whether the Dietary Approaches to Stop Hypertension (DASH) dietary pattern is associated with other blood pressure (BP) variables, beyond mean systolic blood pressure (SBP) and diastolic blood pressure (DBP). OBJECTIVES: The study aimed to study the associations between the DASH dietary pattern and daytime and nighttime mean BPs and BP variance independent of the mean (VIM). METHODS: A sample of 324 Chinese adults aged ≥ 60 y who were not on BP-lowering medications were included in the analysis. The DASH score was calculated using data collected by a validated FFQ. The 24-h ambulatory BP was measured and the mean and VIM SBP and DBP were calculated for both the daytime (06:00-21:59) and nighttime periods (22:00-05:59). Multivariable linear models were constructed to assess associations between the DASH dietary pattern and daytime and nighttime BP outcomes, adjusting for sociodemographic factors, lifestyle, BMI, and hypertension (clinic SBP ≥ 140 mm Hg or DBP ≥ 90 mm Hg), and sleep parameters (only for nighttime BP outcomes). An interaction term between DASH score and hypertension status was added to explore the potential differential association in normotensive and hypertensive individuals. RESULTS: Every 1-unit increase in the DASH score was associated with a 0.18-unit (95% CI: -0.34, -0.01 unit) and a 0.22-unit (95% CI: -0.36, -0.09 unit) decrease in nighttime VIM SBP and nighttime VIM DBP, respectively. DASH score was not associated with any daytime BP outcomes, nighttime mean SBP, or nighttime mean DBP. A significant interaction (DASH score × hypertension status) was detected for VIM SBP (P-interaction = 0.04), indicating a differential association between DASH score and nighttime VIM SBP by hypertension status. CONCLUSIONS: Independently of sleep parameters and other factors, the DASH dietary pattern is associated with lower nighttime BP variability in elderly adults.

Genome-wide analysis of the bZIP gene lineage in apple and functional analysis of MhABF in Malus halliana.

MAIN CONCLUSION: 51 MdbZIP genes were identified from the apple genome by bioinformatics methods. MhABF-OE improved tolerance to saline-alkali stress in Arabidopsis, indicating it is involved in positive regulation of saline-alkali stress response. Saline-alkali stress is a major abiotic stress limiting plant growth all over the world. Members of the bZIP family play an important role in regulating gene expression in response to many kinds of biotic and abiotic stress, including salt stress. According to the transcriptome data, 51 MdbZIP genes responding to saline-alkali stress were identified in apple genome, and their gene structures, conserved protein motifs, phylogenetic analysis, chromosome localization, and promoter cis-acting elements were analyzed. Based on transcriptome data analysis, a MdbZIP family gene (MD15G1081800), which was highly expressed under stress, was selected to isolate and named as MhABF. Expression profile analysis by quantitative real-time PCR confirmed that the expression of MhABF in the leaves of Malus halliana was 10.6-fold higher than that of the control (0 days) after 2 days of stress. Then an MhABF gene was isolated from apple rootstock M. halliana. CaMV35S promoter drived MhABF gene expression vector was constructed to infect Arabidopsis with Agrobacterium-mediated infection. And overexpression MhABF gene plants were obtained. Compared with wild type, transgenic plants grew better under saline-alkali stress and the MhABF-OE lines showed higher chlorophyll content, POD, SOD and CAT activity, which indicated that they had strong resistance to stress. These results indicate that MhABF plays an important role in plant resistance to saline-alkali stress, which lays a foundation for further study on the functions in apple.

Risk stratification for mortality in cardiovascular disease survivors: A survival conditional inference tree analysis.

BACKGROUND AND AIMS: Efficient analysis strategies for complex network with cardiovascular disease (CVD) risk stratification remain lacking. We sought to identify an optimized model to study CVD prognosis using survival conditional inference tree (SCTREE), a machine-learning method. METHODS AND RESULTS: We identified 5379 new onset CVD from 2006 (baseline) to May, 2017 in the Kailuan I study including 101,510 participants (the training dataset). The second cohort composing 1,287 CVD survivors was used to validate the algorithm (the Kailuan II study, n = 57,511). All variables (e.g., age, sex, family history of CVD, metabolic risk factors, renal function indexes, heart rate, atrial fibrillation, and high sensitivity C-reactive protein) were measured at baseline and biennially during the follow-up period. Up to December 2017, we documented 1,104 deaths after CVD in the Kailuan I study and 170 deaths in the Kailuan II study. Older age, hyperglycemia and proteinuria were identified by the SCTREE as main predictors of post-CVD mortality. CVD survivors in the high risk group (presence of 2-3 of these top risk factors), had higher mortality risk in the training dataset (hazard ratio (HR): 5.41; 95% confidence Interval (CI): 4.49-6.52) and in the validation dataset (HR: 6.04; 95%CI: 3.59-10.2), than those in the lowest risk group (presence of 0-1 of these factors). CONCLUSION: Older age, hyperglycemia and proteinuria were the main predictors of post-CVD mortality. TRIAL REGISTRATION: ChiCTR-TNRC-11001489.

Alcohol consumption and risk of cardiovascular disease, cancer and mortality: a prospective cohort study.

BACKGROUND: Studies regarding whether light to moderate alcohol consumption is associated with a lower risk of cardiovascular diseases (CVD) have generated mixed results. Further, few studies have examined the potential impact of alcohol consumption on diverse disease outcomes simultaneously. We aimed to prospectively study the dose-response association between alcohol consumption and risk of CVD, cancer, and mortality. METHODS: This study included 83,732 adult Chinese participants, free of CVD and cancer at baseline. Participants were categorized into 6 groups based on self-report alcohol consumption: 0, 1-25, 26-150, 151-350, 351-750, and > 750 g alcohol/wk. Incident cases of CVD, cancers, and mortality were confirmed by medical records. Hazard ratios (HRs) for the composite risk of these three outcomes, and each individual outcome, were calculated using Cox proportional hazard model. RESULTS: During a median follow-up of 10.0 years, there were 6411 incident cases of CVD, 2947 cancers and 6646 deaths. We observed a J-shaped relation between alcohol intake and risk of CVD, cancer, and mortality, with the lowest risk at 25 g/wk., which is equivalent to ~ 2 servings/wk. Compared to consuming 1-25 g/wk., the adjusted HR for composite outcomes was 1.38 (95% confidence interval (CI):1.29-1.49) for non-drinker, 1.15 (95% CI: 1.04-1.27) for 26-150 g/wk., 1.22 (95% CI: 1.10-1.34) for 151-350 g/wk., 1.33 (95% CI: 1.21-1.46) for 351-750 g/wk., and 1.57 (95% CI: 1.30-1.90) for > 750 g/wk., after adjusting for age, sex, lifestyle, social economic status, and medication use. CONCLUSIONS: Light alcohol consumption at ~ 25 g/wk was associated with lower risk of CVD, cancer, and mortality than none or higher consumption in Chinese adults.

A basic/helix-loop-helix transcription factor controls leaf shape by regulating auxin signaling in apple.

Climate-driven phenological change across local spatial gradients leads to leaf shape variation. At higher elevations, leaves of broadleaf species tend to become narrower, but the underlying molecular mechanism is largely unknown. In this study, a series of morphometric analyses and biochemical assays, combined with functional identification in apple, were performed. We show that the decrease in apple leaf width with increasing altitude is controlled by a basic/helix-loop-helix transcription factor (bHLH TF), MdbHLH3. The MdbHLH3-overexpressing lines have a lower transcript abundance of MdPIN1 encoding an auxin efflux carrier but a higher transcript abundance of MdGH3-2 encoding a putative auxin amido conjugate synthase, resulting in a lower free auxin concentration; feeding the transgenic leaves with exogenous auxin partially restores leaf width. MdbHLH3 transcriptionally suppresses and activates MdPIN1 and MdGH3-2, respectively, by specifically binding to their promoters. This alters auxin homeostasis and transport, consequently leading to changes in leaf shape. These findings suggest that the bHLH TF MdbHLH3 directly modulates auxin signaling in controlling leaf shape in response to local spatial gradients in apple.

Fast Quenching the Burst of Host Salicylic Acid Is Common in Early Strawberry/Colletotrichum fructicola Interaction.

The fungus Colletotrichum fructicola (a species of C. gloeosporioides complex) causes devastating anthracnose in strawberry. Like other species of the genus Colletotrichum, it uses a composite strategy including both the biotrophic and necrotrophic processes for pathogenesis. Host-derived hormones are central regulators of immunity, among which salicylic acid (SA) is the core defense one against biotrophic and hemibiotrophic pathogens. However, the manner and timing of pathogen manipulation of SA are largely elusive in strawberry. To achieve better understanding of the early challenges that SA-mediated defense experiences during strawberry/C. fructicola interaction, dynamic changes of SA levels were followed through the high-performance liquid chromatography method. A very early burst of free SA at 1 h postinoculation (hpi) followed by a fast quenching during the next 12 h was noticed, although rhythm variations were present in two hosts. Transcriptional characterization of genes related to SA pathway in two varieties on C. fructicola inoculation revealed that these genes were differentially expressed, although they were all induced at different time points. At the same time, three types of genes encoding homologous effectors interfering with SA accumulation were found to be first inhibited but sequentially activated during the first 24 hpi. Furthermore, subcellular localization analysis suggests that CfShy1 is a weapon of C. fructicola for strawberry invasion. Based on these results, we propose that the infection strategy that C. fructicola utilizes on strawberry is specialized, which is implemented through the optimized expression of a specific set of effector genes. Transcriptional characterization of host genes supports that de novo SA biosynthesis and the free SA release from methyl salicylate might contribute equally to the intricate control of SA homeostasis in strawberry. C. fructicola manipulation of SA-dependent resistance in strawberry might be closely related to multihormonal interplay among SA, jasmonic acid, abscisic acid, and cytokinin.

Prospective Study of Fasting Blood Glucose and Intracerebral Hemorrhagic Risk.

BACKGROUND AND PURPOSE: Although diabetes mellitus is an established independent risk factor for ischemic stroke, the association between fasting blood glucose and intracerebral hemorrhage (ICH) is limited and inconsistent. The objective of the current study was to examine the potential impact of long-term fasting blood glucose concentration on subsequent risk of ICH. METHODS: This prospective study included 96 110 participants of the Kailuan study, living in Kailuan community, Tangshan city, China, who were free of cardiovascular diseases and cancer at baseline (2006). Fasting blood glucose concentration was measured in 2006, 2008, 2010, and 2012. Updated cumulative average fasting blood glucose concentration was used as primary exposure of the current study. Incident ICH from 2006 to 2015 was confirmed by review of medical records. RESULTS: During 817 531 person-years of follow-up, we identified 755 incident ICH cases. The nadir risk of ICH was observed at fasting blood glucose concentration of 5.3 mmol/L. The adjusted hazard ratios and their 95% confidence intervals (CIs) of ICH were 1.59 (95% CI, 1.26-2.02) for diabetes mellitus or fasting blood glucose ≥7.00 mmol/L, 1.31 (95% CI, 1.02-1.69) for impaired fasting blood glucose (fasting blood glucose, 6.10-6.99 mmol/L), 0.98 (95% CI, 0.78-1.22) for fasting blood glucose 5.60 to 6.09 mmol/L, and 2.04 (95% CI, 1.23-3.38) for hypoglycemia (fasting blood glucose, <4.00 mmol/L), comparing with normal fasting blood glucose 4.00 to 5.59 mmol/L. The results persisted after excluding individuals who used hypoglycemic, aspirin, antihypertensive agents, or anticoagulants, and those with intracerebral hemorrhagic cases occurred in the first 2 years of follow-up. CONCLUSIONS: In this large community-based cohort, low (<4.0 mmol/L) and high (≥6.1 mmol/L) fasting blood glucose concentrations were associated with higher risk of incident ICH, relative to fasting blood glucose concentrations of 4.00 to 6.09 mmol/L.

Transcriptional and physiological analyses of short-term Iron deficiency response in apple seedlings provide insight into the regulation involved in photosynthesis.

BACKGROUND: Iron (Fe) is an essential micronutrient for plants. Utilization of Fe deficiency-tolerant rootstock is an effective strategy to prevent Fe deficiency problems in fruit trees production. Malus halliana is an apple rootstock that is resistant to Fe deficiency; however, few molecular studies have been conducted on M. halliana. RESULTS: To evaluate short-term molecular response of M. halliana leaves under Fe deficiency condition, RNA sequencing (RNA-Seq) analyses were conducted at 0 (T1), 0.5 (T2) and 3 d (T3) after Fe-deficiency stress, and the timepoints were determined with a preliminary physiological experiment. In all, 6907, 5328, and 3593 differentially expressed genes (DEGs) were identified in pairs of T2 vs. T1, T3 vs. T1, and T3 vs. T2. Several of the enriched DEGs were related to heme binding, Fe ion binding, thylakoid membranes, photosystem II, photosynthesis-antenna protein, porphyrin and chlorophyll metabolism and carotenoid biosynthesis under Fe deficiency, which suggests that Fe deficiency mainly affects the photosynthesis of M. halliana. Additionally, we found that Fe deficiency induced significant down-regulation in genes involved in photosynthesis at T2 when seedlings were treated with Fe-deficient solution for 0.5 d, indicating that there was a rapid response of M. halliana to Fe deficiency. A strong up-regulation of photosynthesis genes was detected at T3, which suggested that M. halliana was able to recover photosynthesis after prolonged Fe starvation. A similar expression pattern was found in pigment regulation, including genes for coding chlorophyllide a oxygenase (CAO), ß-carotene hydroxylase (ß-OHase), zeaxanthin epoxidase (ZEP) and 9-cis-epoxycarotenoid dioxygenase (NCED). Our results suggest that pigment regulation plays an important role in the Fe deficiency response. In addition, we verified sixteen genes related to photosynthesis-antenna protein, porphyrin and chlorophyll metabolism and carotenoid biosynthesis pathways using quantitative real-time PCR (qRT-PCR) to ensure the accuracy of transcriptome data. Photosynthetic parameters, Chl fluorescence parameters and the activity of Chlase were also determined. CONCLUSIONS: This study broadly characterizes a molecular mechanism in which pigment and photosynthesis-related regulations play indispensable roles in the response of M. halliana to short-term Fe deficiency and provides a basis for future analyses of the key genes involved in the tolerance of Fe deficiency.

Transcriptome analysis in Malus halliana roots in response to iron deficiency reveals insight into sugar regulation.

Iron (Fe) deficiency is a frequent nutritional problem limiting apple production in calcareous soils. The utilization of rootstock that is resistant to Fe deficiency is an effective way to solve this problem. Malus halliana is an Fe deficiency-tolerant rootstock; however, few molecular studies have been conducted on M. halliana. In the present work, a transcriptome analysis was combined with qRT-PCR and sugar measurements to investigate Fe deficiency responses in M. halliana roots at 0 h (T1), 12 h (T2) and 72 h (T3) after Fe deficiency stress. Total of 2473, 661, and 776 differentially expressed genes (DEGs) were identified in the pairs of T2 vs. T1, T3 vs. T1, and T3 vs. T2, respectively. Several DEGs were enriched in the photosynthesis, glycolysis and gluconeogenesis, tyrosine metabolism and fatty acid degradation pathways. The glycolysis and photosynthesis pathways were upregulated under Fe deficiency. In this experiment, sucrose accumulated in Fe-deficient roots and leaves. However, the glucose content significantly decreased in the roots, while the fructose content significantly decreased in the leaves. Additionally, 15 genes related to glycolysis and sugar synthesis and sugar transport were selected to validate the accuracy of the transcriptome data by qRT-PCR. Overall, these results indicated that sugar synthesis and metabolism in the roots were affected by Fe deficiency. Sugar regulation is a way by which M. halliana responds to Fe deficiency stress.