OsMAPK6 phosphorylation and CLG1 ubiquitylation of GW6a non-additively enhance rice grain size through stabilization of the substrate.

The chromatin modifier GRAIN WEIGHT 6a (GW6a) enhances rice grain size and yield. However, little is known about its gene network determining grain size. Here, we report that MITOGEN-ACTIVED PROTEIN KINASE 6 (OsMAPK6) and E3 ligase CHANG LI GENG 1 (CLG1) interact with and target GW6a for phosphorylation and ubiquitylation, respectively. Unexpectedly, however, in vitro and in vivo assays reveal that both of the two post-translational modifications stabilize GW6a. Furthermore, we uncover two major GW6a phosphorylation sites (serine142 and threonine186) targeted by OsMAPK6 serving an important role in modulating grain size. In addition, our genetic and molecular results suggest that the OsMAPK6-GW6a and CLG1-GW6a axes are crucial and operate in a non-additive manner to control grain size. Overall, our findings identify a previously unknown mechanism by which phosphorylation and ubiquitylation non-additively stabilize GW6a to enhance grain size, and reveal correlations and interactions of these posttranslational modifications during rice grain development.

Control of grain size in rice by TGW3 phosphorylation of OsIAA10 through potentiation of OsIAA10-OsARF4-mediated auxin signaling.

Grain size is a key component of grain yield and quality in crops. Several core players of auxin signaling have been revealed to modulate grain size; however, to date, few genetically defined pathways have been reported, and whether phosphorylation could boost degradation of Aux/IAA proteins is uncertain. Here, we show that TGW3 (also called OsGSK5) interacts with and phosphorylates OsIAA10. Phosphorylation of OsIAA10 facilitates its interaction with OsTIR1 and subsequent destabilization, but this modification hinders its interaction with OsARF4. Our genetic and molecular evidence identifies an OsTIR1-OsIAA10-OsARF4 axis as key for grain size control. In addition, physiological and molecular studies suggest that TGW3 mediates the brassinosteroid response, the effect of which can be relayed through the regulatory axis. Collectively, these findings define a auxin signaling pathway to regulate grain size, in which phosphorylation of OsIAA10 enhances its proteolysis and potentiates OsIAA10-OsARF4-mediated auxin signaling.

CaSBP11 Participates in the Defense Response of Pepper to Phytophthora capsici through Regulating the Expression of Defense-Related Genes.

Squamosa promoter binding protein (SBP)-box genes are plant-specific transcription factors involved in plant growth and development, morphogenesis and biotic and abiotic stress responses. However, these genes have been understudied in pepper, especially with respect to defense responses to Phytophthora capsici infection. CaSBP11 is a SBP-box family gene in pepper that was identified in our previous research. Silencing CaSBP11 enhanced the defense response of pepper plants to Phytophthora capsici. Without treatment, the expression of defense-related genes (CaBPR1, CaPO1, CaSAR8.2 and CaDEF1) increased in CaSBP11-silenced plants. However, the expression levels of these genes were inhibited under transient CaSBP11 expression. CaSBP11 overexpression in transgenic Nicotiana benthamiana decreased defense responses, while in Arabidopsis, it induced or inhibited the expression of genes in the salicylic acid and jasmonic acid signaling pathways. CaSBP11 overexpression in sid2-2 mutants induced AtNPR1, AtNPR3, AtNPR4, AtPAD4, AtEDS1, AtEDS5, AtMPK4 and AtNDR1 expression, while AtSARD1 and AtTGA6 expression was inhibited. CaSBP11 overexpression in coi1-21 and coi1-22 mutants, respectively, inhibited AtPDF1.2 expression and induced AtPR1 expression. These results indicate CaSBP11 has a negative regulatory effect on defense responses to Phytophthora capsici. Moreover, it may participate in the defense response of pepper to Phytophthora capsici by regulating defense-related genes and the salicylic and jasmonic acid-mediated disease resistance signaling pathways.

Identification of Pepper CaSBP08 Gene in Defense Response Against Phytophthora capsici Infection.

Little information is available on the role of Squamosa promoter binding protein (SBP)-box genes in pepper plants. This family of genes is known to have transcription characteristics specific to plants and to regulate plant growth, development, stress responses, and signal transduction. To investigate their specific effects in pepper (Capsicum annuum), we screened pepper SBP-box family genes (CaSBP genes) for Phytophthora capsici (P. capsici) resistance genes using virus-induced gene silencing. CaSBP08, CaSBP11, CaSBP12, and CaSBP13, which are associated with plant defense responses against P. capsici, were obtained from among fifteen identified CaSBP genes. The function of CaSBP08 was identified in pepper defense response against P. capsici infection in particular. CaSBP08 protein was localized to the nucleus. Silencing of CaSBP08 enhanced resistance to P. capsici infection. Following P. capsici inoculation, the malondialdehyde content, peroxidase activity, and disease index percentage of the CaSBP08-silenced plants decreased compared to the control. Additionally, the expression levels of other defense-related genes, especially those of CaBPR1 and CaSAR8.2, were more strongly induced in CaSBP08-silenced plants than in the control. However, CaSBP08 overexpression in Nicotiana benthamiana enhanced susceptibility to P. capsici infection. This work provides a foundation for the further research on the role of CaSBP genes in plant defense responses against P. capsici infection.

Transcription Factor CaSBP12 Negatively Regulates Salt Stress Tolerance in Pepper (Capsicum annuum L.).

SBP-box (Squamosa-promoter binding protein) genes are a type of plant-specific transcription factor and play important roles in plant growth, signal transduction, and stress response. However, little is known about the role of pepper SBP-box transcription factor genes in response to abiotic stress. Here, one of the pepper SBP-box gene, CaSBP12, was selected and isolated from pepper genome database in our previous study. The CaSBP12 gene was induced under salt stress. Silencing the CaSBP12 gene enhanced pepper plant tolerance to salt stress. The accumulation of reactive oxygen species (ROS) of the detached leaves of CaSBP12-silenced plants was significantly lower than that of control plants. Besides, the Na+, malondialdehyde content, and conductivity were significantly increased in control plants than that in the CaSBP12-silenced plants. In addition, the CaSBP12 over-expressed Nicotiana benthamiana plants were more susceptible to salt stress with higher damage severity index percentage and accumulation of ROS as compared to the wild-type. These results indicated that CaSBP12 negatively regulates salt stress tolerance in pepper may relate to ROS signaling cascades.

A small heat shock protein CaHsp25.9 positively regulates heat, salt, and drought stress tolerance in pepper (Capsicum annuum L.).

Extreme environmental conditions seriously affect crop growth and development, resulting in a decrease in crop yield and quality. However, small heat shock proteins (Hsp20s) play an important role in helping plants to avoid these negative impacts. In this study, we identified the expression pattern of the CaHsp25.9 gene in a thermo-tolerance pepper line R9 and thermo-sensitive line B6. The transcription of CaHsp25.9 was strongly induced by heat stress in both R9 and B6. The expression of CaHsp25.9 was induced by salt and drought stress in R9. Additionally, the CaHsp25.9 protein was localized in the cell membrane and cytoplasm. When silencing the CaHsp25.9 gene in the R9 line, the accumulation of malonaldehyde (MDA), relative electrolytic leakage, hydrogen peroxide, superoxide anion were increased, while total chlorophyll decreased under heat, salt, and drought stress. Over-expression of CaHsp25.9 in Arabidopsis resulted in decreased MDA, while proline, superoxide dismutase activity, germination, and root length increased under heat, salt, and drought stress. However, peroxidase activity was higher in drought stress but lower in heat and salt stress in transgenic Arabidopsis compared to the wild type (WT). Furthermore, the transcription of stress related genes was more highly induced in transgenic lines than WT. Our results indicated that CaHsp25.9 confers heat, salt, and drought stress tolerance to plants by reducing the accumulation of reactive oxygen species, enhancing the activity of antioxidant enzymes, and regulating the expression of stress-related genes. Therefore, these results may provide insight into plant adaption mechanisms developed in variable environments.

A Novel Transcription Factor CaSBP12 Gene Negatively Regulates the Defense Response against Phytophthora capsici in Pepper (Capsicum annuum L.).

SBP-box (Squamosa-promoter binding protein) genes are a type of plant-specific transcription factor and play important roles in plant growth, signal transduction and stress response. However, little is known about the SBP-box genes in pepper (CaSBP), especially in the process of Phytophthora capsici infection. In this study, a novel gene (CaSBP12) was selected from the CaSBP gene family, which was isolated from the pepper genome database in our previous study. The CaSBP12 gene was located in the nucleus of the cell and its silencing in the pepper plant enhanced the defense response against Phytophthora capsici infection. After inoculation with Phytophthora capsici, the root activity of the CaSBP12-silenced plants is compared to control plants, while malondialdehyde (MDA) content is compared viceversa. Additionally, the expression of defense related genes (CaPO1, CaSAR8.2, CaBPR1, and CaDEF1) in the silenced plants were induced to different degrees and the peak of CaSAR8.2 and CaBPR1 were higher than that of CaDEF1. The CaSBP12 over-expressed Nicotiana benthamiana plants were more susceptible to Phytophthora capsici infection with higher EC (electrical conductivity) and MDA contents as compared to the wild-type. The relative expression of defense related genes (NbDEF, NbNPR1, NbPR1a, and NbPR1b) in transgenic and wild-type Nicotiana benthamiana plants were induced, especially the NbPR1a and NbPR1b. In conclusion, these results indicate that CaSBP12 gene negative regulates the defense response against Phytophthora capsici infection which suggests their potentially significant role in plant defense. To our knowledge, this is the first report on CaSBP gene which negative regulate defense response.

[The Application of Gold-immunochromatographic Test Strip Reader in Serum Antibody Detection in Echinococcosis].

One hundred and fifty-nine serum samples from hydatid disease patients and 80 serum samples from patients with other liver diseases were detected by gold-immunochromatographic assay, and read by naked eyes and the gold-immunochromatographic test strip reader. The sensitivity, specificity, and accuracy of eye-based method was 92.4% (147/159), 85.0% (68/80), and 89.9% (215/239), which was lower than that of the reader detection (95.6%, 93.7%, 95.0%, respectively). While, its false negative rate (7.5%, 12/159) and false positive rate (15.0%, 12/80) was higher than that of the reader detection (4.4% and 6.3%, respectively).

[Prevalence of human and ovine hepatic hydatid disease diagnosed by ultrasound in Hobukesar Mongolian Autonomous County of Xinjiang].

OBJECTIVE: To investigate the prevalence of human and ovine hepatic hydatid disease in Hobukesar Mongolian Autonomous County of Xinjiang (HMACX)and to evaluate the related strategies for prevention and control of the disease. METHODS: A prevalence screening method was used to screen local residents and sheep for hydatid disease in HMACX. Based on B ultrasound images, the screening programs on people and sheep in different sites were carried and the findings were comparatively analyzed. RESULTS: Findings of B ultrasound images through screening program among human beings showed that the positive rates of hydatid diseases 4.4% (23/521), of cystic echinococcosis and alveolar echinococcosis as 4.0% (21/521) and 0.8% (4/521)respectively. The infection rate on sheep was 3.8% (7/180). The positive rates of human and ovine hepatic hydatid disease in Township Chagangule were higher than in other areas. There was no significant statistical difference noticed on human positive rates between Township Chagangule and other areas. Statistically, significant difference for positive rate in ovine was seen between Township Chagangule and Township Bayinaowa(χ(2) = 4.8259, P = 0.0280). As intermediate host of hydatid disease, the infection rate in sheep was higher than that in human beings at Township Chagangule. CONCLUSION: HMACX remained a highly endemic area for human and ovine hydatid disease.

[Effects of Chinese herbal medicine Qianggu Capsule on patients with rheumatoid arthritis-induced osteoporosis: a report of 82 cases].

BACKGROUND: Rheumatoid arthritis (RA) is a kind of chronic autoimmune disease and osteoporosis is one of its complications. OBJECTIVE: To explore the effects of Qianggu Capsule, a compound traditional Chinese herbal medicine, on bone mineral density (BMD) and osteoporosis in patients with RA. DESIGN, SETTING, PARTICIPANTS AND INTERVENTION: Eighty-two patients with rheumatoid arthritis and osteoporosis, who were treated in Shanghai Guanghua Hospital of Integrated Traditional Chinese and Western Medicine from January 2010 to December 2011, were divided into treatment group (42 cases) and control group (40 cases). The patients in the treatment group were administered with Qianggu Capsule and two disease-modifying antirheumatic drugs. The patients in the control group were administered with two common-used antirheumatic drugs. The course of treatment was 6 months. MAIN OUTCOME MEASURES: Blood levels of alkaline phosphatase (ALP), calcium, phosphorus, C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) were determined before and after the treatment. BMD in the lumbar spine, femur and the left distal radius were also examined before and after the treatment. RESULTS: The ALP level, a bone metabolic parameter, was significantly increased in patients of the treatment group after treatment compared with before treatment. BMD values in the lumbar spine, femur and the radius were higher after treatment than before treatment (P<0.05). There were no changes in ALP level and BMD in the patients of the control group after the treatment when compared with before treatment. CONCLUSION: Treatment with Qianggu Capsule can increase BMD of RA patients, and then ameliorate their osteoporosis.

[Fine root production and turnover in Pinus massoniana plantation in Three Gorges Reservoir area of China].

By the methods of sequential soil cores and buried bags, an investigation was conducted to study the seasonal dynamics of fine roots in a 20-year-old Pinus massoniana plantation in Three Gorges Reservoir Area from March to December 2011, with the annual production and turnover rate of the fine roots calculated. In the plantation, the annual mean biomass of <2 mm fine roots was 146.98 g x m(-2) x a(-1), in which, the living root biomass (102.92 g x m(-2) x a(-1)) was far greater than that of the dead root biomass (44.06 g x m(-2) x a(-1)). Among the fine roots with different sizes, <1 mm fine roots had an obvious seasonal dynamics in their biomass, showing a unimodal curve in the sampling period. The annual production and turnover rate of <2 mm fine roots were 104. 12 g x m(-2) x 1(-1) and 1.05 a(-1), respectively, in which, the annual production of <1 mm and 1-2 mm fine roots was 58.35 and 45.77 g x m(-2) x a(-1), and the turnover rate was 1.41 and 0.69 a(-1), respectively.

[Synthesis and luminescence properties of Y10W2O21:Eu nanophosphor].

In the present paper, a novel nanophosphor, Y10W2O21:Eu was synthesized through a simple and low-cost method: co-precipitation. The results of XRD show that resultant samples are Orthorhombic phase and primitive lattice. The average crystallographic sizes could be confirmed to be approximately 80 nm by means of the Scherrer formula, which are in good agreement with the particles sizes exhibited by SEM. In the excitation spectra of Y10W2O21:Eu nanophosphor, by monitoring 610 nm, the charge transfer bands (CTB) of O--Eu and O--W, centering at around 270 and 307 nm, can be observed, respectively. The spectral lines shape and locations of excitation peak corresponding to 4f-->4f transitions are similar in all samples. But the relative intensity ratios between O--Eu and O--W CTB excitation peak increase with the Eu3+ doping concentration increasing. The characteristic red emission at around 610 nm of Eu3+ was also observed, ascribed to the (5)D0--(7)>F2 transition of Eu3+, and the optimal doping concentration is 20 mol%. Finally, the transition intensity parameters omega(lamda = 2,4) and the quantum efficiencies of (5)D0 level of Eu3+ were calculated according to Judd-Ofelt theory. The results indicate that Eu(3+5)D0-->(7)F2 610 nm red luminescence can be effectively excited by 394 nm near-UV light and 464 nm blue light in Y10W2O21 host, which is similar to the familiar Eu3+ doped tungstate (e.g., Gd2(WO4)3, CaWO4). Therefore, the Y10W2O21:Eu red nanophosphors may have a potential application for white LED.

[Risk factors on human cystic echinococcosis in Hobukesar Mongolian Autonomous County in Xinjiang].

OBJECTIVE: To study the risk factors of human cystic echinococcosis (CE) in Hobukesar Mongolian Autonomous County of Xinjiang (HMACX) and to discuss the related strategies for prevention and control. METHODS: A randomized sampling method was used to screen local residents for human CE in HMACX. CE related risk factors including ethnicity, age, sex, occupation and personal status on hygiene etc. were analyzed under multi-factor logistic regression. RESULTS: The prevalence rates of CE and alveolar echinococcosis (AE) were 3.7% (23/627) and 0.16% (1/627) respectively, with the seropositive rate as 12.4% (76/613). The main risk factors that significantly associated with CE were age (OR = 7.6, 95%CI: 2.481 - 23.579) and slaughtering livestock in the households (OR = 3.2, 95%CI: 1.297 - 7.809). Herdsmen had the highest prevalence of CE in all of the occupations in this study. CONCLUSION: HMACX had been a highly endemic area for human CE, with age and family slaughtering-livestock-behavior appeared to be the main possible risk factors.

[Characteristics of soil seed banks in backwater area of Three Gorges Reservoir water-level-fluctuating zone at initial stage of river-flooding].

A germination test was made to study the characteristics of soil seed banks in the backwater area (including the flooded, non-flooded, and control areas of secondary bush and abandoned farmland) of Three Gorges Reservoir water-level-fluctuating zone. There existed significant differences in the soil seed banks between secondary bush and abandoned farmland, with an average seed density being 6991 +/- 954 seed per m2 and 26193 +/- 6928 seed per m2, respectively. Flooded area had the lowest seed density, while non-flooded area had the highest one. The seed density decreased with soil depth. A total of 118 species belonging to 45 families and 97 genera were detected in the soil seed banks of secondary bush and abandoned farmland, most of which were annual and perennial herbage species, belonging to Asteraceae, Poaceae, Scrophulariaceae, and Cruciferae. Among the 118 species, there were 34 species (occupying 28.8%) whose individuals accounted for less than 0.01% of the total. In the soil seed banks of secondary bush and abandoned farmland, the species number was similar, species diversity index and evenness index were relatively high, but the dominant species differed greatly, and the ecological dominance was relatively low. The species diversity in non-flooded area and the ecological dominance in flooded area were the highest, and the similarity index between the flooded and non-flooded areas was the highest.

[Study on the variation of lymphocytes and cytokines in patients with echinococcosis granulosus].

OBJECTIVE: To investigate the variation of lymphocytes and cytokines in patients with cystic echinococcosis (CE). METHODS: 80 CE patients who were diagnosed for the first time (60 of Han and 20 of Uygur nationality), and 37 patients who were to accept the second surgical operation(24 Han and 13 Uygur nationality) were included in the study. The peripheral lymphocytes of patients before operation were analyzed by flow cytometry (FCM) to detect T and B lymphocytes, NK cells bearing surface markers, as well as Thl cytokine IFN-gamma and Th2 cytokine IL-4 in the cytoplasm of lymphocytes. 179 healthy persons served as control. RESULTS: In the group of Han patients who were diagnosed for the first time, the percentage of total T cells(CD3+) was lower than the control (P<0.05), while among patients accepting the second operation, the ratio of total T cells showed no difference to the control. For the helper T cells (CD3+/CD4+), NK cells (CD3+/CD16,56+) and B cells (CD3+/CD19+), their ratio were significantly lower in both groups than the control (P<0.01), but their cytotoxic T cells (CD3+/CD8+) were higher than the control. In Uygur patients diagnosed for the first time, B cell ratio was lower than that in control (P<0.05). The NK cell level in both groups of patients was lower than control (P<0.01 and P<0.05 respectively). The level of ThO and Th1 showed no statistical difference among the three groups (P>0.05). The Th2 level was significantly higher in the first diagnosed patients than control (P<0.01), but no statistical difference between the group with the second operation and the other two groups. CONCLUSION: The immune status of echinococcosis patients is inhibited and the level of Th1 and Th2 shows difference in the first and second operation groups.

[Expression of Smad4 in leukemia cells].

Loss of transforming growth factor (TGF)-beta signaling has been implicated in malignant transformation of various tissues. Smad4 plays a central role in the signal transduction of TGF-beta. Deletion or mutation of Smad4 has been described in a number of cancers. This study was aimed to investigate a potential role of Smad4 in leukemia including its expression and location in blast cells. The mononuclear cells were separated from bone marrow of leukemia patients. The samples, blast cells of which were more than 90% in mononuclear cells, were selected. The expression and location of Smad4 protein were analyzed by immunohistochemistry methods. The results showed that the Smad4 protein located mainly in nucleus, part of this protein located in cytoplasma, the expressions of Smad4 were not detected in 6 out of 9 ALL patients, in 7 out of 24 AML patients and in 1 out of 2 CML patients; these leukemia patients, in whose cells the expression of Smad4 was not detected, included one L1 and one L3, four L2, one M0, one M1, two M2a, one M3a, one M4b, one M6 and one CML. In conclusion, the Smad4 protein was mainly in nucleus, the deletion or functional change of Smad4 may related with the pathogenesis of human AML.

[Field trial on rapid detection of echinococcosis by dot immunogold filtration assay (DIGFA) with whole blood sample].

OBJECTIVE: To establish a rapid, simple and reliable assay with samples of whole blood for diagnosis and epidemiological study on hydatidosis. METHODS: The dot immunogold filtration assay kit was developed and potato agglutinin was applied to blot blood quickly. RESULTS: Among 1 678 persons from prevalent area, the positive rate of DIGFA was 8.469 while that of image examination was 3.04%. Both DIGFA and image technique showed positive results in 43 cases. 8 cases with positive image but negative DIGFA were followed up for 16 months, which turned out that 3 cases with necrotic hydatid cysts, 2 cases with calcified hydatid cysts and 2 cases with benign hepatic cysts. 99 cases with positive DIGFA but negative image were also followed up for 16 months, 3 pulmonary hydatid cases were confirmed. Among 38 cases proved by operation and histopathology, the positive rate of DIGFA was 89.5%. 52 samples from non-prevalent area all showed negative DIGFA. Another 40 non-hydatidosis cases (10 samples of hepatic hemangioma, 10 of non-parasitic cysts of liver, 10 of primary hepatic carcinoma, 6 of pulmonary tuberculosis, 4 of lung cancer) also showed negative DIGFA. 190 samples were selected randomly and detected blindly by DIGFA with whole blood, DIGFA with serum and ELISA with serum to evaluate their diagnostic effect with no statistical difference (P>0.05). CONCLUSION: The DIGFA kit is rapid, simple and reliable in epidemiological study of hydatid disease, with an advantage of using whole blood sample instead of serum.

[RT-PCR detecting NUP98-HOX fusion gene in leukemia].

To investigate whether there are NUP98-HOXA, NUP98-HOXB, NUP98-HOXC, NUP98-HOXD fusion genes in leukemia patients in Xinjiang, cellular total RNA was extracted from the bone marrow mononuclear cells, the formaldehyde-agarose gel electrophoresis was used to judge whether RNA was intact, the 17 RT-PCR primers were designed to amplify the predicted fusion junctions and 412 bp GAPDH was used as an internal control, NUP98-HOXA fusion genes were amplified by nested-PCR following reverse transcription. One-step PCR was performed to amplify the other predicted fusion genes. The results showed that RNA was proved to be intact and expression of GAPDH was found in every sample. However, no predicted fusion transcripts were detected in leukemia patients. In conclusion, no NUP98-HOX fusion genes were detected in the samples from Xinjiang.

Identification of a gene engineering antibody against cystic echinococcosis in liver.

OBJECTIVE: To identify a gene engineering antibody against cystic echinococcosis in liver. METHODS: A single chain of variable fragment of human antibodies (ScFvs) was selected from the library by using affinity selection technique with the recombinant antigen on solid surface. The positive clones were demonstrated by ELISA and their DNA sequences were also determined. RESULTS: The DNA sequence data showed that the antibody gene is composed of 768 bp. In addition, a specific combination capacity with recombinant Echinococcus granulosus antigen B (r-EgB) was demonstrated by ELISA. CONCLUSION: The obtained gene engineering antibody against r-EgB may have potential implications in immunological treatment and drug targeting delivery.