The molecular mechanism of WRINKLED1 transcription factor regulating oil accumulation in developing seeds of castor bean.

The transcription factor WRINKLED1 (WRI1), a member of AP2 gene family that contain typical AP2 domains, has been considered as a master regulator regulating oil biosynthesis in oilseeds. However, the regulatory mechanism of RcWRI1 in regulating oil accumulation during seed development has not been clearly addressed. Castor bean (Ricinus communis) is one of the most important non-edible oil crops and its seed oils are rich in hydroxy fatty acids, widely applied in industry. In this study, based on castor bean reference genome, three RcWRIs genes (RcWRI1, RcWRI2 and RcWRI3) were identified and the expressed association of RcWRI1 with oil accumulation were determined. Heterologous transformation of RcWRI1 significantly increased oil content in tobacco leaf, confirming that RcWRI1 activate lipid biosynthesis pathway. Using DNA Affinity Purification sequencing (DAP-seq) technology, we confirmed RcWRI1 binding with Transcription Start Site of genes and identified 7961 WRI1-binding candidate genes. Functionally, these identified genes were mainly involved in diverse metabolism pathways (including lipid biosynthesis). Three cis-elements AW-box ([CnTnG](n)7[CG]) and AW-boxes like ([GnAnC](n)6[GC]/[GnAnC](n)7[G]) bound with RcWRI1 were identified. Co-expression network analysis of RcWRI1 further found that RcWRI1 might be widely involved in biosynthesis of storage materials during seed development. In particular, yeast one hybrid experiments found that both AP2 domains within RcWRI1 were required in binding targeted genes. These results not only provide new evidence to understand the regulatory mechanism of RcWRI1 in regulation of oil accumulation during castor bean seed development, but also give candidate gene resource for subsequent genetic improvement toward increasing oil content in oilseed crops.

Engineering an oilseed crop for hyper-accumulation of carotenoids in the seeds without using a traditional marker gene.

KEY MESSAGE: Ketocarotenoids were synthesized successfully in Camelina sativa seeds by genetic modification without using a traditional selection marker genes. This method provided an interesting tool for metabolic engineering of seed crops. Camelina sativa (L.) Crantz is an important oil crop with many excellent agronomic traits. This model oil plant has been exploited to accumulate value-added bioproducts using genetic manipulation that depends on antibiotic- or herbicide-based selection marker genes (SMG), one of the major concerns for genetically modified foods. Here we reported metabolic engineering of C. sativa to synthesize red ketocarotenoids that could serve as a reporter to visualize transgenic events without using a traditional SMG. Overexpression of a non-native ß-carotene ketolase gene coupled with three other carotenogenous genes (phytoene synthase, ß-carotene hydroxylase, and Orange) in C. sativa resulted in production of red seeds that were visibly distinguishable from the normal yellow ones. Constitutive expression of the transgenes led to delayed plant development and seed germination. In contrast, seed-specific transformants demonstrated normal growth and seed germination despite the accumulation of up to 70-fold the level of carotenoids in the seeds compared to the controls, including significant amounts of astaxanthin and keto-lutein. As a result, the transgenic seed oils exhibited much higher antioxidant activity. No significant changes were found in the profiles of fatty acids between transgenic and control seeds. This study provided an interesting tool for metabolic engineering of seed crops without using a disputed SMG.

Genome-wide analysis of the B3 transcription factors reveals that RcABI3/VP1 subfamily plays important roles in seed development and oil storage in castor bean (Ricinus communis).

The B3 transcription factors (TFs) in plants play vital roles in numerous biological processes. Although B3 genes have been broadly identified in many plants, little is known about their potential functions in mediating seed development and material accumulation. Castor bean (Ricinus communis) is a non-edible oilseed crop considered an ideal model system for seed biology research. Here, we identified a total of 61 B3 genes in the castor bean genome, which can be classified into five subfamilies, including ABI3/VP1, HSI, ARF, RAV and REM. The expression profiles revealed that RcABI3/VP1 subfamily genes are significantly up-regulated in the middle and later stages of seed development, indicating that these genes may be associated with the accumulation of storage oils. Furthermore, through yeast one-hybrid and tobacco transient expression assays, we detected that ABI3/VP1 subfamily member RcLEC2 directly regulates the transcription of RcOleosin2, which encodes an oil-body structural protein. This finding suggests that RcLEC2, as a seed-specific TF, may be involved in the regulation of storage materials accumulation. This study provides novel insights into the potential roles and molecular basis of B3 family proteins in seed development and material accumulation.

Anti-Inflammatory and Anti-Oxidative Effects of Isorhamnetin for Protection Against Lung Injury in a Rat Model of Heatstroke in a Dry-Heat Environment.

BACKGROUND Isorhamnetin is a natural flavonoid compound with anti-inflammatory and antioxidant properties. However, its roles in alleviating lung injury associated with heatstroke remain unclear. Therefore, this study aimed to evaluate the protective effects of different isorhamnetin doses on lung injury in heatstroke rat models exposed to a dry-heat environment. MATERIAL AND METHODS Fifty Sprague-Dawley rats were randomly divided into 5 groups: normal control (0.9% saline), heatstroke (0.5% CMCNa), and isorhamnetin (25, 50, and 100 mg/kg) groups; treatments were administered by gavage daily for 7 days. All rats, except those in the control group, were exposed to a dry-heat environment (41±1°C, 10±2% relative humidity) for 150 min to induce heatstroke. Pathological changes, ultrastructure, edema, inflammation, and oxidative stress in the lungs were assessed. RESULTS Compared with the heatstroke group, rats treated with 100 mg/kg isorhamnetin showed amelioration of histopathological and ultrastructural changes in the lungs; decreased lung injury scores (P<0.05) and wet/dry weight ratios (P<0.01); lower levels of phospho-nuclear factor-kappaB (P<0.05), high-mobility group box 1 (P<0.01), tumor necrosis factor-alpha (P<0.01), interleukin (IL)-1ß (P<0.01), and IL-6 (P<0.01); lower malondialdehyde contents (P<0.01); and higher superoxide dismutase (P<0.01) and catalase activities (P<0.05). CONCLUSIONS In a dry-heat environment, isorhamnetin protected against lung injury in heatstroke rat models via anti-inflammatory and anti-oxidative mechanisms.

[Cadmium tolerance and enrichment characteristics of Eulaliopsis binata].

A pot experiment was conducted to study the cadmium (Cd) tolerance and enrichment characteristics of Eulaliopsis binata. Applying low concentration Cd (5 mg x kg(-1)) could boost the growth and physiological vigor of E. binata, with the twelve indexes such as biomass and net photosynthetic rate, etc. increased by 1.0% -15.5%, compared with CK; while applying high concentration Cd (> 5 mg x kg(-1)) had negative effects on the growth of E. binata. Even though the E. binata still finished its physiological circle under the application of 100 mg Cd x kg(-1), the biomass and net photosynthetic rate were decreased by 27.0% and 25.6%, respectively, in comparison with CK. The Cd content in the organs of E. binata increased greatly with increasing concentration of applied Cd. The Cd content in roots was from 350 mg x kg(-1) to 500 mg x kg(-1), and that in stems and leaves was from 15 mg x kg(-1) to 35 mg x kg(-1). When the concentration of applied Cd was < 50 mg x kg(-1), the Cd uptake by E. binata increased with increasing concentration of applied Cd. The Cd content in stems and leaves amounted 15.7% - 38.4% of the total uptake, and the ratio of the Cd content in stems and leaves to that in roots was up to 0.62. All the results indicated that E. binata had stronger capability in tolerating, enriching, and transporting Cd, being a potential Cd hyperaccumulator.

[Effect of health service and its influence factors in flood disaster areas].

OBJECTIVE: To explore the effect of health service and its influential factors in flood disaster areas. METHODS: Fifty-five towns were sampled randomly from Dongting Lake area suffering from flood in 1998. The health service level, effect, and its influence factors were investigated retrospectively. RESULTS: The incidence rate of notifiable infectious diseases was 11.7 per thousand, prevalence rate of chronic disease was 51.2 per thousand, infant mortality rate was 43.1 per thousand, neonatal mortality rate was 10.2 per thousand, and the total mortality rate was 554.3/100,000 in Dongting Lake area. The health investment level was significantly associated with the incidence rate of notifiable infectious diseases, the infant mortality rate, and total mortality rate. Duration of flood and income per capita were important factors for the effect of health investment. CONCLUSION: Increasing health investment and residents' income in the flood disaster area, shortening the duration of flood would play positive role in residents' health in the flood disaster area.

[Years of potential life of residents among different flood areas in Hunan province].

OBJECTIVE: To determine the damage of different types of floods on the residents health. METHODS: The methods of standard mortality rate (SMR) and years of potential life lost (YPLL) were used to analyze the death of sample residents from flood areas in Dongting Lake in Hunan province. RESULTS: The order of death causes in the soaked area, the collapsed area and the non-flood area was the same. But the mortality rates of residents injury, poisoning and malignant neoplasm diseases in the soaked area and the collapsed area were higher than those of non-flood area. The resident standard rates of years of potential life lost (SYPLL) in the soaked area and the collapsed area were higher than that of the controls, especially in the age group of 30 to 45. The flood-attributed SYPLL in the male was higher than that of the female. CONCLUSION: Flood actually affected the health of residents. The more serious the flood is, the worse the effect is. It is very important to decrease the resident mortality rate of the injury, poisoning and malignant neoplasm, and to pay attention to protect people of 30 to 45 years old in flood areas.

The molecular phylogeny of Alpinia (Zingiberaceae): a complex and polyphyletic genus of gingers.

Alpinia is the largest, most widespread, and most taxonomically complex genus in the Zingiberaceae with 230 species occurring throughout tropical and subtropical Asia. Species of Alpinia often predominate in the understory of forests, while others are important ornamentals and medicinals. Investigations of the evolutionary relationships of a subset of species of Alpinia using DNA sequence-based methods specifically test the monophyly of the genus and the validity of the previous classifications. Seventy-two species of Alpinia, 27 non-Alpinia species in the subfamily Alpinioideae, eight species in the subfamily Zingiberoideae, one species in the subfamily Tamijioideae, and three species in the outgroup genus Siphonochilus (Siphonochiloideae) were sequenced for the plastid matK region and the nuclear internal transcribed spacer (ITS) loci. Parsimony analyses of both individual and combined data sets identified six polyphyletic clades containing species of Alpinia distributed across the tribe Alpinieae. These results were supported by a Bayesian analysis of the combined data set. Except in a few specific cases, these monophyletic groupings of species do not correspond with either Schumann's (1904) or Smith's (1990) classification of the genus. Here we build on previous molecular analyses of the Alpinioideae and propose the next steps necessary to recognize new generic boundaries in the Alpinieae.

[A research on the public health index related to the comprehensive assessment on floods].

OBJECTIVE: To develop a public health index related to the comprehensive assessment on the impact caused by floods. METHODS: A Analytic hierarchy process (AHP) theory was used to establish the initial evaluation system on the impact of floods. Modified-Delphi process was used to screen and determine the indicators and their weights while synthetical scored method was used to establish the comprehensive assessment model. Percentile was used to differentiate the degree of floods. Finally, analysis of variance (ANOVA) and correlation analysis were used to test the differentiability of the model for different degree of floods and the independence of these indicators. RESULTS: The model of comprehensive assessment on the impact of floods was set up, including six first-ranking indicators and twenty-four sub-indicators. The values of comprehensive assessment were divided into five grades by the percentiles. Verified results indicated that there were significant difference among the five grades (F = 76.11, P < 0.01) and all indicators were independent. CONCLUSION: An index of comprehensive assessment on the impact of floods was established, which could be used to evaluate the impact of floods and to differentiate the degree of flood, which seemed to have the characteristic of reliability, comprehensiveness and practicability.

[A study on the health status of residents affected by flood disasters].

OBJECTIVE: To study the immediate and long-term effects of disasters caused by floods on residents health status. METHODS: Stratified sampling by ranks of flood disaster occurred in 1996 and 1998, flood disaster areas and control areas were carried out. A retrospective study was also carried out to study all diseases involved during 1996 - 1999. RESULTS: The incident rates of acute infectious disease in flooding areas in 1996 and 1998 were both higher than those of non-flooding areas (863.181/100 000 and 736.591/100 000, respectively). But there was no different between the incident rate of the first years in flooding areas and that of non-flooding areas. The prevalence rates of 8 kinds of chronic diseases related to circulatory system, nervous system, digestive system, injury and poisonous diseases in flooding areas were also higher than that in the non-flooding areas. The highest incidence rates of most diseases were in the mountainous flooding areas, followed by areas collapsed by flooding, and the lowest were seen in soakedareas by floods. The incidence rates of intestinal infectious diseases and respiratory infectious diseases were lower in areas where prevention and control measures were weak. CONCLUSION: Flood could lead to the increase of incidence rates both on acute infectious diseases and non-infectious diseases. Interventions on non-infectious diseases should also be enforced to stop the epidemics when preventing and controlling acute infectious disease.

[Study on hospitalization expenses of flood disaster areas' residents of Dongting Lake in Hunan province in 1998].

OBJECTIVE: To study the expenses of hospitalization among the population in the flood disaster areas of Dongting Lake in Hunan province in 1998. METHODS: Descriptive epidemiologic study were conducted to analyze hospitalization expenses of the residents of 55 villages in flood disaster areas in 1998; single factors analysis and logarithmic linear regression analysis were carried out to explore influencing factors about hospitalization expenses of the residents. RESULTS: The hospitalization rate was 4.59% with an average hospitalization expenses of 667.42 Yuan in the flood disaster areas' residents of Dongting Lake in 1998. Compared with populations without suffering from flood, hospitalization rate and the average hospitalization expenses of flood disaster Areas' residents of Dongting Lake in 1998 were higher and had significant difference. The average hospitalization expenses in 1998 was affected by flood types, family income, gender, age, literacy, occupation, outcome after leaving the hospital and hospital ranks. CONCLUSION: These results implied that the flood disease aggravated inhabitants' burden of disease in Dongting Lake areas; the factors influencing the average hospitalization expenses were multiple, and synthetic measures should be taken in the prevention and control of flood disaster.