Seven new aconitine-type diterpenoid alkaloids from the roots of Aconitum apetalum.

Seven new aconitine-type C19-diterpenoid alkaloids, apetaldines K-Q (1-7), were isolated from the roots of Aconitum apetalum (Huth) B. Fedtsch. Their structures were established on the basis of extensive spectroscopic analyses (HRESIMS, 1D and 2D NMR). Among them, compounds 1 and 2 possess a unique 2-(E)-(2-methylbut-2-enamido)benzoate moiety at the C-18 position. Furthermore, cytotoxic activities of these diterpenoid alkaloids were also evaluated.

Neuroelectrophysiological alteration associated with cognitive flexibility after 24 h sleep deprivation in adolescents.

The cognitive neural mechanisms by which sleep deprivation affects cognitive flexibility are poorly understood. Therefore, the study investigated the neuroelectrophysiological basis of the effect of 24 h sleep deprivation on cognitive flexibility in adolescents. 72 participants (36 females, mean age ± SD=20.46 ± 2.385 years old) participated in the study and were randomly assigned to the sleep deprivation group and control group. They were instructed to complete a task switch paradigm, during which participants' behavioral and electroencephalographic data were recorded. Behaviorally, there were significant between-group differences in accuracy. The results of event-related potential showed that the P2, N2 and P3 components had significant group effects or interaction effects. At the time-frequency level, there were statistically significant differences between the delta and theta bands. These results suggested that 24 h sleep deprivation affected problem-solving effectiveness rather than efficiency, mainly because it systematically impaired cognitive processing associated with cognitive flexibility.

Electrophysiological indexes of ingroup bias in a group Stroop task: Evidence from an event-related potential study.

Although cognitive system assigns higher attentional resources to ingroup information than outgroup information, but it is unclear whether the ingroup bias can be measured by the processes that are related to allocation of attentional resources to ingroup information. Thus, a group Stroop task was developed to study the issues combining with event-related potential (ERP) technique in this study. Specifically, 34 subjects (17 female, mean age = 20.76 ±â€¯1.26) were firstly divided into blue or red group (17 subjects for each group); then they were asked to categorize four words of Stroop task into "our team" or "other team" based on the ink color (blue/red) of the words whose meaning were also red/blue. The behavioral results showed that outgroup ink color processing was interfered by ingroup word meaning, but the ingroup ink color processing was less/not interfered by outgroup word meaning. The ERP results showed that the amplitude of frontal N100 was enhanced when more attentional resources were automatically captured by ingroup information in early stage than outgroup information; P2/N2 amplitude was reduced or enhanced when outgroup information processing was interfered by ingroup information; enhanced P3b amplitude reflected that attention could be more easily allocated to ingroup information than outgroup information based on target. This study implied a novel direction to study the neural basis of ingroup bias by investigating the roles of ingroup bias in assigning attentional resources to group information.

Longitudinal analysis of the mediating role of self-perception of aging in the relationship between frailty and intrinsic capacity: A cross-lagged mediation model.

OBJECTIVES: The causal relationship between intrinsic capacity and frailty in older adults, as well as the underlying temporal mechanisms, remained poorly understood. The study aimed to investigate the causal association between intrinsic capacity and frailty while exploring the potential mediating role of self-perception of aging. MEASUREMENTS: A survey was conducted with a sample of 429 participants who completed measures of intrinsic capacity, self-perception of aging, and frailty at baseline and were followed for one year. The relationships between these variables were assessed using an autoregressive cross-lagged model. RESULTS: The study found reciprocal associations between intrinsic capacity and frailty (p < 0.01). Furthermore, the results indicated that self-perception of aging partially mediated the effect of frailty at baseline (T1) on intrinsic capacity at one-year follow-up (T2) (ß = -0.02, confidence interval: [-0.055, -0.004]). However, the reverse causation was not observed. CONCLUSIONS AND IMPLICATIONS: This study demonstrates a bidirectional causal relationship between intrinsic capacity and frailty in older adults. Self-perception of aging plays a significant mediating role in this relationship. Older adults with a worse level of frailty should be made aware of the potentially vicious cycle related to self-perception of aging, which can negatively affect their intrinsic capacity. Maintaining a positive self-perception of aging may help preserve physical and psychological reserves, maintain intrinsic capacity, and slow the decline of frailty.

UBE3A and MCM6 synergistically regulate the proliferation and migration of lung adenocarcinoma cells.

Lung cancer is a leading cause of mortality worldwide and shows substantial clinical and biomolecular heterogeneity. Currently, specific therapeutic strategies are lacking, so effective drug targets are urgently needed. E6AP/UBE3A is a multifaceted ubiquitin ligase that controls various signaling pathways implicated in neurological diseases and various cancers; however, its role in lung cancer is incompletely understood. Here, MCM6 was identified as an interacting partner of E6AP using the yeast two-hybrid assay. MCM2 and MCM4 were then shown to interact with E6AP. E6AP knockout enhanced the ubiquitination of MCM2/4/6, suggesting that E6AP was not the E3 ubiquitin ligase for these three MCM proteins. Ablation of E6AP inhibited proliferation and migration, but had no significant effect on apoptosis in A549 and H1975 cells, and proliferation and migration inhibition was also observed in MCM6 knockdown cells. Furthermore, ablation of MCM6 and E6AP synergistically suppressed the proliferation and migration of A549 and H1975 cells. To verify the above findings in vivo, we established tumor models in nude mice and identified that the tumorigenicity of human lung adenocarcinoma (LUAD) cells was synergistically regulated by MCM6 and E6AP. Moreover, the expression levels of MCM6 and E6AP were higher in LUAD tissues than in adjacent tissues. Furthermore, the expression levels of MCM6 and E6AP were positively correlated in human LUAD samples. Thus, our study suggests that the interaction of E6AP and MCM proteins plays an important role in the progression of LUAD, which might offer potential therapeutic targets for cancer treatment.

TRIM25-mediated ubiquitination of G3BP1 regulates the proliferation and migration of human neuroblastoma cells.

Neuroblastoma is one of the most severe malignant tumors and accounts for substantial cancer-related mortality in children. Ras-GTPase-activating protein SH3 domain-binding protein 1 (G3BP1) is highly expressed in various cancers and acts as an important biomarker of poor prognosis. The ablation of G3BP1 inhibited the proliferation and migration of human SHSY5Y cells. Because of its important role in neuroblastoma, the regulation of G3BP1 protein homeostasis was probed. TRIM25, which belongs to the tripartite motif (TRIM) family of proteins, was identified as an interacting partner for G3BP1 using the yeast two-hybrid (Y2H) method. TRIM25 mediates the ubiquitination of G3BP1 at multiple sites and stabilizes its protein level. Then, our study found that TRIM25 knockdown also inhibited the proliferation and migration of neuroblastoma cells. The TRIM25 and G3BP1 double knockdown SHSY5Y cell line was generated, and double knockdown cells exhibited lower proliferation and migration ability than cells with only TRIM25 or G3BP1 knockdown. Further study demonstrated that TRIM25 promotes the proliferation and migration of neuroblastoma cells in a G3BP1-dependent manner. Tumor xenograft assays indicated that the ablation of TRIM25 and G3BP1 synergistically suppressed the tumorigenicity of neuroblastoma cells in nude mice, and TRIM25 promoted the tumorigenicity of G3BP1 intact SHSY5Y cells but not G3BP1 knockout cells. Thus, TRIM25 and G3BP1, two oncogenic genes, are suggested as potential therapeutic targets for neuroblastoma.

Antipsychotics-induced improvement of cool executive function in individuals living with schizophrenia.

Cool executive dysfunction is a crucial feature in people living with schizophrenia which is related to cognition impairment and the severity of the clinical symptoms. Based on electroencephalogram (EEG), our current study explored the change of brain network under the cool executive tasks in individuals living with schizophrenia before and after atypical antipsychotic treatment (before_TR vs. after_TR). 21 patients with schizophrenia and 24 healthy controls completed the cool executive tasks, involving the Tower of Hanoi Task (THT) and Trail-Marking Test A-B (TMT A-B). The results of this study uncovered that the reaction time of the after_TR group was much shorter than that of the before_TR group in the TMT-A and TMT-B. And the after_TR group showed fewer error numbers in the TMT-B than those of the before_TR group. Concerning the functional network, stronger DMN-like linkages were found in the before_TR group compared to the control group. Finally, we adopted a multiple linear regression model based on the change network properties to predict the patient's PANSS change ratio. Together, the findings deepened our understanding of cool executive function in individuals living with schizophrenia and might provide physiological information to reliably predict the clinical efficacy of schizophrenia after atypical antipsychotic treatment.

Association between positive control in self-perceptions of aging and motoric cognitive risk syndrome among Chinese community-dwelling older adults: a cross-sectional study.

BACKGROUND: Self-perceptions of aging (SPA) are important psychosocial factors that lead to a wide range of outcomes including dementia. However, the relationships between positive SPA and motoric cognitive risk syndrome (MCR) which is a predementia syndrome are still unknown. This study aimed to reveal the associations of positive control and aging awareness of SPA with the risk of MCR and its components. METHODS: A cross-sectional design was conducted among 1137 Chinese community-dwelling older adults. Positive control and aging awareness were defined by two dimensions of SPA (Positive control and Timeline chronic). MCR was determined according to definition. Multivariable logistic regression was used to examine the associations. RESULTS: The overall prevalence of MCR was 11.5% (mean age = 71.62 ± 5.22). After adjusting for depression, anxiety, and cognitive function, positive control was associated with reduced risk of MCR (OR = 0.624, 95% CI 0.402-0.969, P = 0.036), subjective cognitive complaints (SCC) (OR = 0.687, 95% CI 0.492-0.959, P = 0.027), and gait speed (GS) (OR = 0.377, 95% CI 0.197-0.720, P = 0.003), respectively. Aging awareness was merely related to increased risk of MCR (OR = 1.386, 95% CI 1.062-1.810, P = 0.016). CONCLUSIONS: This study highlights the crucial associations of positive control and aging awareness with MCR and its components. Our results emphasize that positive belief in control and adaptive aging awareness might be promising targets for preventing MCR.

MKRN1/2 serve as tumor suppressors in renal clear cell carcinoma by regulating the expression of p53.

BACKGROUND: Kidney renal clear cell carcinoma (KIRC) belongs to renal cell carcinoma which is a very aggressive malignant tumor with poor prognosis and high mortality. The MKRN family includes three members MKRN1, MKRN2 and MKRN3, which are closely related to cancers, and have been involved in many studies. OBJECTIVE: This study aimed to explore the roles of MKRN family in KIRC. METHODS: The expression of MKRNs was analyzed using the UALCAN database, prognostic analysis was performed with the GEPIA2 and Kaplan-Meier Plotter database, and correlation analysis was assessed by GEPIA2. The CCK-8 and colony formation assay were performed to detect cell proliferation, wound healing assays were performed to detect cell migration, cell cycles were detected by flow cytometry analysis, GST pull-down and co-immunoprecipitation assays were performed to detect the interaction of proteins, and the expression of MKRNs, p53 and other proteins were detect by immunoblotting analysis or quantitative PCR (qPCR). RESULTS: MKRN1 and MKRN2 were lowly expressed in KIRC samples compared to the corresponding normal tissues, and KIRC patients with high levels of MKRN1 and MKRN2 showed higher overall survival (OS) and disease free survival (DFS) rates. The overexpression of MKRN1 and MKRN2 inhibited the proliferation of human KIRC cells by arresting the cell cycles, but shows little effect on cells migration. The expression of MKRN1 and MKRN2 are correlated, and MKRN1 directly interacts with MKRN2. Moreover, both MKRN1 and MKRN2 were closely correlated with the expression of TP53 in KIRC tumor, and promoted the expression of p53 both at protein and mRNA levels. CONCLUSIONS: Our study suggests that MKRN1 and MKRN2 serve as tumor suppressors in KIRC, and act as promising therapeutic targets for KIRC treatment.

Cueing effect of attention among nurses with different anxiety levels: an EEG study.

The attention to cueing among nurses with anxiety affects their nursing quality seriously. Nevertheless, the neural mechanism of attention under anxiety among nurses has not been revealed. In this study, we utilized the event-related potential (ERP) and functional brain networks to investigate the neural mechanism of the cueing attention differences between anxiety and non-anxiety nurse groups (AG-20 nurses; NAG-20 nurses) in the spatial cueing task. The results revealed that in the invalid cues (144 trials), longer reaction times, larger P2 amplitudes, and more linkages between the right frontal and parietal areas were found in AG compared to NAG. In the valid cues (288 trials), there were no significant behavioral and neural differences between the two groups. The AG in the invalid cues showed slower response times, larger P2 and N5 amplitudes, and denser linkages originating from the occipital cortex than those in the valid cues. The convolutional neural network was trained for discriminating between the anxiety nurses and the normal ones, with the average accuracy being 0.76. The findings provided a potential physiological biomarker to predict the anxiety group who need to give more psychological attention. Nurse leaders maybe get more information for offering solutions to retain mental health among nurses.

The association between serum orexin-A levels and sleep quality in pregnant women.

OBJECTIVE/BACKGROUND: Orexin has been shown to regulate the sleep-wake cycle, and it may play a major role in the pathogenesis of sleep disorders; however, its role in sleep disorders in pregnant women remains unclear. We aimed to assess the relationship between serum orexin-A (OXA) levels and sleep quality in pregnant women. PATIENTS/METHODS: This study comprised 214 enrolled pregnant women (poor sleep quality, n = 125; no poor sleep quality, n = 89). We assessed participants' sleep quality and depression and anxiety levels. OXA levels were measured using enzyme-linked immunosorbent assay. RESULTS: Women in the poor sleep quality group showed higher serum OXA levels (0.33[0.3] vs. 0.27[0.11], P < 0.001) than those in the no poor sleep quality group. Binary regression analysis showed that the higher the OXA levels (odds ratio [OR] 1.385, 95% CI [confidence interval] 1.160-1.655) and Zung Self-Rating Anxiety Scale scores (OR 1.073, 95% CI 1.009-1.140), the greater the risk of sleep quality in pregnant women. First-trimester OXA levels differed significantly from those in the second and third trimesters (P < 0.05). CONCLUSION: Serum OXA levels were higher in pregnant women with poor sleep quality than in those without poor sleep quality. OXA levels were also higher in the second and third trimesters than in the first trimester.

Dynamical changes of land use/land cover and their impacts on ecological quality during China's reform periods: A case study of Quanzhou city, China.

The rapid growth of China's economy has greatly accelerated the process of urbanization during China's reform periods. Urbanization has significantly caused land use and land cover (LULC) changes and thus has impacts on the local climate and ecosystem. This study chooses Quanzhou, a fast-developing city of southeast China, as an example to detect and quantify the LULC and ecological changes from 1989 to 2018 by using the remotely sensed technique. The LULC of Quanzhou was derived from the four Landsat images taken in 1989, 1999, 2007 and 2018, and the land-use-degree ratio index and land-use-change method were used to estimate the change of land use. The remote sensing based ecological index (RSEI) was used to detect the ecological changes of the city. The built-up land expansion intensity and annual built-up land expansion rate were carried out for seven districts of Quanzhou. The results show that the urban area of Quanzhou has drastically grown by 192.99 km2 at the expense of forest, water, and cropland land during the 1989~2018 period. Moreover, the built-up land of seven districts had expanded at the average rate of 0.027~0.154 per year and the built-up expansion intensity was higher than 0.59. The average RSEI value of Quanzhou city dropped from 0.78 in 1989 to 0.34 in 2018, which suggested an overall decline in ecological quality. The proportion of areas with an RSEI rating good decreased from 30.84% to 11.52% while the proportion of areas with rating bad increased from 4.73% to 19.11% during the past 29 years. This study has shown the built-up land expansion intensity is negatively correlated with the ecological quality change, and the increase in built-up land can greatly accelerate the decline of the ecological quality. Government policies play a profound impact on land use changes, urbanization and eco-environment changes. Therefore, the policy decision-makers should take enough action and consider integrating the concept of ecology to enable the healthy and sustainable development of the city.

Genome-wide identification of SHMT family genes in cucumber (Cucumis sativus L.) and functional analyses of CsSHMTs in response to hormones and abiotic stresses.

Serine hydroxymethyltransferase (SHMT) is a pyridoxal phosphate-dependent enzyme that plays crucial roles in the photorespiration and one-carbon metabolism of plants. In the present research, we conducted a systematic analysis of the SHMT gene family in cucumber (Cucumis sativus L). Results show that a total of 6 SHMT members were identified from the cucumber genome database. CsSHMT1 and CsSHMT2 participate in a fragment duplication event, indicating that CsSHMTs may complete the expansion of family members through fragment duplication. Gene structure analysis found that the number of exons of CsSHMTs ranges from 4 to 15. Members with the same number of exons are classified into the same class in the phylogenetic analysis. Each class reflects its subcellular distribution. Expression and function analysis reveals that CsSHMTs express in a variety of plant tissues, indicating that SHMT gene expression pattern is not organ-specific. qRT-PCR analysis found that CsSHMT3 and CsSHMT5 positively respond to abscisic acid (ABA), and CsSHMT2-6 are induced by indole-3-acetic acid (IAA) and methyl jasmonate (MeJA). Abiotic stress analysis shows that CsSHMT3 is significantly induced by drought and salt stress. These results may provide useful information for further function and evolution analysis of cucumber SHMT genes. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03378-x.

The chloroplast genome of Rosa rugosa × Rosa sertata (Rosaceae): genome structure and comparative analysis.

Rosa rugosa × Rosa sertata, which belongs to the family Rosaceae, is one of the native oil-bearing roses in China. Most research has focused on its essential oil components and medicinal values. However, there have been few studies about its chloroplast genome. In this study, the whole chloroplast genome of R. rugosa × R. sertata was sequenced, analyzed, and compared to other genus Rosa species. The chloroplast genome of R. rugosa × R. sertata is a circular structure and 157,120 bp in length. The large single copy and small single copy is 86,173 bp and 18,743 bp in size, respectively, and the inverted repeats are 26,102 bp in size. The GC content of the whole genome is 37.96%, while those of regions of LSC, SSC, and IR are 35.20%, 31.18%, and 42.73%, respectively. There are 130 different genes annotated in this chloroplast genome, including 84 protein coding genes, 37 tRNA genes, 8 rRNA genes, and 1 pseudogene. Phylogenetic analysis of 19 species revealed that R. rugosa × R. sertata belong to the Sect. Cinnamomeae. Overall, this study, providing genomic resources of R. rugosa × R. sertata, will be beneficial for species identification and biological research.

Two New C18 -Diterpenoid Alkaloids from Aconitum leucostomum Worosch.

Two new lappaconitine-type C18 -diterpenoid alkaloids, named as leucostosines C (1) and D (2), together with six known compounds (3-8), were isolated from the roots of Aconitum leucostomum Worosch. Their structures were elucidated by various spectroscopic analyses, including IR, HR-ESI-MS, NMR spectra and X-ray experiments. Leucostosine C is the first diterpenoid alkaloid bearing the 7-amino group. The isolated compounds were tested for the acetylcholinesterase (AChE) inhibitory effect and neuroprotective activity, none of them showed significant activities.

Self-reference processing of fat-face and sick-face in individuals with different disgust sensitivity: Evidence from behavioral and neuroelectrophysiology.

Self-reference effect has been widely discussed. Previous scholars believed that self-related information can be processed faster is due to the positive attribute of self-concept which speeds up self-related information processing. When self-related information is given negative attributes, the self-reference effect will be weakened. In this study, fat and sick, two kinds of stimuli associated with disgust characteristics, were added to self- and other-faces. We found that disease stimuli, which are closely related to survival threats, eliminated the self-reference effect while the obesity stimuli only weakened the self-reference effect to a certain extent. Event-related potential (ERP) analysis demonstrated that sick-faces have a greater amplitude than standard-faces in the EPN and LPP components. We believe that this may be due to the urgency of the disease threat, which leads to the selective attention to the disease threat in the early perception stage and the allocation of more attention resources for rapid response in the later stage. In addition, we found that disgust sensitivity specifically maintains individuals' self-referential effects by dissociating individuals from others in disease contexts. These results further support the behavioral immune function of disgust as a gatekeeper of the self in potentially contaminated environments. In conclusion, our study showed that in the face of survival threat, the self-reference effect is eliminated, and disgust tried to slow down this elimination effect to protect self. This study extends the behavioral immunity theory to some extent and further deepens the understanding of the relationship between disgust and self.

The Number of MRGPRX2-Expressing Cells Is Increased in Skin Lesions of Patients With Indolent Systemic Mastocytosis, But Is Not Linked to Symptom Severity.

Background: Recently, the expression of the mast cell (MC) receptor Mas-related G protein-coupled receptor X2 (MRGPRX2) has been detected in lesional skin of adult patients with cutaneous mastocytosis. As of yet, little is known about the clinical relevance of MRGPRX2 and its agonists in patients with mastocytosis, including indolent systemic mastocytosis (ISM). Methods: MRGPRX2 and MRGPRX2 agonists, cortistatin (CST), and major basic protein (MBP) were analyzed in lesional and non-lesional skin of patients with ISM and skin of healthy controls by immunohistochemistry. Co-localization of MRGPRX2 and MRGPRX2-mRNA with the MC marker tryptase was assessed by immunofluorescence microscopy and in situ hybridization, respectively. We assessed clinical, demographic, and laboratory data, including mastocytosis activity score (MAS), serum tryptase, and KIT D816V allele burden. Results: The number of MRGPRX2-expressing (MRGPRX2+) cells, MRGPRX2-mRNA+ MCs, and CST-expressing (CST+) and MBP-expressing (MBP+) cells was significantly higher in lesional skin as compared to non-lesional skin and/or skin of healthy controls (all p < 0.05). Increased numbers of MRGPRX2+ cells, MRGPRX2-mRNA+ MCs, and CST+ and MBP+ cells were not associated with clinical and laboratory features of ISM, including disease burden, symptom severity, evidence of anaphylaxis, and tryptase levels. Conclusions: Skin lesions of patients with ISM showed high numbers of MRGPRX2+ cells, although they were not linked to symptom severity. Clinical relevance of the MRGPRX2-mediated pathway of MC activation in ISM remains unclear and should be investigated in further studies.

Depression and Cognition Mediate the Effect of Self-Perceptions of Aging Over Frailty Among Older Adults Living in the Community in China.

Objectives: The aims of the study were first to investigate the association between self-perceptions of aging and frailty and second to determine whether self-perceptions of aging affects frailty via depressive symptoms and cognitive status among older adults living in the community in China. Methods: Among 850 older adults who participated in this cross-sectional study, 822 older adults made valid responses to Tilburg Frailty Indicator, Brief Aging Perceptions Questionnaire, UCLA loneliness scale-8, Mini-Mental State Examination, and Patient Health Questionnaire-9 between March to December 2019. The possible pathways of self-perceptions of aging affecting frailty were analyzed based on the structural equation modeling analysis. Results: A total of 21.53% of older adults reported frailty. Correlation analyses showed that higher degrees of frailty were related to greater loneliness, more depressive symptoms, more negative self-perceptions of aging, worse locomotive function, and cognitive status (r = 0.267, r = 0.440, r = 0.481, r = 0.451, r = -0.337; p < 0.001). Multiple regression analysis showed that loneliness, depressive symptoms, self-perceptions of aging, locomotive function, and cognitive status were the five factors to be entered the regression equation, and the variance of joint explanation was 46.60%. SPA had a direct effect on frailty (ß = 0.306 and p < 0.001), and SPA indirectly affects frailty by independently affecting depressive symptoms (ß = 0.391, 95% CI [0.027, 0.061], and p < 0.001) or cognitive status (ß = 0.148, 95% CI [0.009, 0.024], and p < 0.001) of older adults. Conclusion: These findings help explain the potential psychological mechanisms through which SPA impacts frailty and may aid community healthcare providers in China in identifying individuals at high risk of frailty. The results suggest that health staff should help older adults improve their perspectives on aging, alleviate or prevent depressive symptoms, and improve cognitive status to delay the progress of frailty and promote healthy aging.

JAZ1-3 and MYC2-1 Synergistically Regulate the Transformation from Completely Mixed Flower Buds to Female Flower Buds in Castanea mollisima.

Chestnut (Castanea mollisima) is an important woody food crop, but its yield has been low in cultivation, mainly due to the problems of fewer female flowers and more male flowers. Therefore, regulating the transition of chestnut flowers and effectively balancing the proportion of male and female to improve the yield are key factor to be solved in production. In this study, the chestnut floral buds in pre- and post-winter were used as materials. The data of metabolites, hormones, and gene expression during flower bud differentiation of chestnut were analyzed by transcriptomics and metabolomics to preliminarily reveal the possible reason of male and female flower bud transformation in pre- and post-winter. The analysis of Differentially Expressed Genes (DEGs) showed that there were 6323 DEGs in the Complete mixed flower bud (CMF) group in pre- and post-winter, of which 3448 genes were up-regulated and 2875 genes were down-regulated. There were 8037 DEGs in the Incomplete mixed flower bud (IMF) in pre- and post-winter, of which 4546 genes were up-regulated and 3491 genes were down-regulated. A total of 726 genes from the two flower buds were enriched into 251 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways in post winter, of which plant hormone signal transduction accounted for 4.13%. The analysis results of differential metabolites showed that the differential metabolites of the two flower buds were mainly concentrated in the secondary metabolic synthesis pathway. The difference of hormone content showed that the content of Gibberellin 9 (GA9) and GA19 in CMF was higher than that in IMF in pre-winter, but the opposite in post-winter. Methyl jasmonate (MeJA) content was only very high in CMF in pre-winter, while Jasmonoyl-(l)-Isoleucine (JA-ILE) showed high content in CMF in post-winter. In post-winter, higher concentration of JA-ILE was positively correlated with the expression of Flowering Locus T (CmFT), and CmFT gene was significantly positively correlated with the expression levels of MYC2-1, MYC2-2 and LFY 3 (LEAFY 3). The higher concentration of JA-ILE was negatively correlated with the transcription level of JAZ1-3. In vitro experiments further verified that Jasmonate-Zim 1-3 (JAZ 1-3) combined with MYC2-1 inhibited the transcription of CmFT gene, while MYC2-1 alone promoted the expression of FT. The results suggested that a higher concentration of GA is conducive to breaking the dormancy of flower buds and promoting the development of male flower buds, while a lower concentration of GA and a higher concentration of JA-ILE are conducive to the differentiation and formation of female flower buds in post-winter, in which JAZ1-3 and MYC2-1 play a key role in the differentiation of female flower buds of chestnut.

An optimized method to obtain high-quality RNA from different tissues in Lilium davidii var. unicolor.

The high quality, yield and purity total RNA samples are essential for molecular experiments. However, harvesting high quality RNA in Lilium davidii var. unicolor is a great challenge due to its polysaccharides, polyphenols and other secondary metabolites. In this study, different RNA extraction methods, namely TRIzol method, the modified TRIzol method, Kit method and cetyltrimethylammonium bromide (CTAB) method were employed to obtain total RNA from different tissues in L. davidii var. unicolor. A Nano drop spectrophotometer and 1% agarose gel electrophoresis were used to detect the RNA quality and integrity. Compared with TRIzol, Kit and CTAB methods, the modified TRIzol method obtained higher RNA concentrations from different tissues and the A260/A280 ratios of RNA samples were ranged from 1.97 to 2.27. Thus, the modified TRIzol method was shown to be the most effective RNA extraction protocol in acquiring RNA with high concentrations. Furthermore, the RNA samples isolated by the modified TRIzol and Kit methods were intact, whereas different degrees of degradation happened within RNA samples isolated by the TRIzol and CTAB methods. In addition, the modified TRIzol method could also isolate high-quality RNA from other edible lily bulbs. Taken together, the modified TRIzol method is an efficient method for total RNA isolation from L. davidii var. unicolor.