Accumulation of specific flavonoids in soybean (Glycine max (L.) Merr.) as a function of the early tripartite symbiosis with arbuscular mycorrhizal fungi and Bradyrhizobium japonicum (Kirchner) Jordan

This study is the first report assessing the effect of soil inoculation on the signalling interaction of Bradyrhizobium japonicum, arbuscular mycorrhizal fungi (AMF) and soybean plants throughout the early stages of colonisation that lead to the tripartite symbiosis. In a study using soil disturbance to produce contrasting indigenous AMF treatments, the flavonoids daidzein, genistein and coumestrol were identified as possible signals for regulating the establishment of the tripartite symbiosis. However, it was unclear whether soil disturbance induced changes in flavonoid root accumulation other than through changing the potential for AMF colonization. In this study, soil treatments comprising all possible combinations of AMF and B. japonicum were established to test whether (1) modifications in root flavonoid accumulation depend on the potential for AMF colonization, and (2) synthesis and accumulation of flavonoids in the roots change over time as a function of the early plant-microbial interactions that lead to the tripartite symbiosis. The study was comprised of two phases. First, maize was grown over 3-week periods to promote the development of the AM fungus Glomus clarum. Second, the interaction between soybean, G. clarum and B. japonicum was evaluated at 6, 10, 14 and 40 days after plant emergence. Root colonization by G. clarum had a positive effect on nodulation 14 days after emergence, producing, 30% more nodules which were 40% heavier than those on roots solely inoculated with B. japonicum. The tripartite symbiosis resulted in 23% more N₂ being fixed than did the simpler symbiosis between soybean and B. japonicum. The presence of both symbionts changed accumulation of flavonoids in roots. Daidzein and coumestrol increased with plant growth. However, development of the tripartite symbiosis caused a decrease in coumestrol; accumulation of daidzein, the most abundant flavonoid, was reduced in the presence of AMF.     

Manganese modulates the responses of nitrogen-supplied and Rhizobium-nodulated Phaseolus vulgaris L. to inoculation with arbuscular mycorrhizal fungi

In Venezuela, low yields of black bean crops are attributed, in part, to the low manganese (Mn) and phosphorus (P) contents in the Quartzipsamment soils where this crop is usually sown. To test this hypothesis, black bean plants were grown in sterilized sand to simulate soil physical properties, were fertilized with increasing Mn concentrations (0.1-20 μM) and inoculated with a commercial mixture of Rhizobium leguminosarum bv phaseoli strains 127K44, 127K89, 127K105 (+Rh), in combination with arbuscular mycorrhizal fungi Scutellospora heterogama and Entrophospora colombiana (+AMF). Non-inoculated plants fertilized with 6 mM NO₃ and 2 mM P served as controls. Plants were harvested at 18, 25, 33, and 40 days after emergence. At all harvests, the greatest growth and highest P and iron (Fe) leaf concentrations occurred in control plants grown in 5 μM Mn. The growth of +AMF plants was promoted at 0.1 μM Mn and inhibited at higher than 1 μM Mn. Whereas, concentrations of 5-10 μM Mn enhanced the growth and the Mn concentrations in leaves of +Rh plants 40 days after emergence. The tripartite symbiosis (+Rh+AMF) decreased growth, nodulation and leaf ureide and chlorophyll concentrations in plants grown in less than 20 μM Mn, imputed to severe ultrastructural alterations in the leaf and nodule tissues. Only +Rh+AMF plants grown in 20 μM Mn were effectively nodulated, AMF colonized and reached the flowering stage, although with diminished growth and low chlorophyll concentrations. Results confirm the high Mn requirement of +Rh plants for growth and nodulation and question the implementation of the tripartite symbiosis to improve yields in early flowering black bean varieties planted in soils deficient in Mn and P.     

苏州农村“三大合作”——中国城乡一体化发展之路

苏州农村"三大合作"是农村社区股份合作社、土地股份合作社和农民专业合作社的统称.苏州"三大合作"较好地解决了农村发展中遇到的各种瓶颈问题--集体资产权利主体不明确、缺乏规模经营、市场地位弱势等,维护了农民在经济社会发展中的基本权利,促进了农村、农业和农民的整体较快发展."三大合作"是对农村生产关系的主动调整,它既体现了社会主义的核心价值观,又确保了市场经济条件下农村社会的现代化进程,是一条具有鲜明中国特色的城乡一体化发展之路.     

农业产业投资基金合作三方三阶段动态博弈分析

基于博弈论视角,构建了农业产业投资基金、农业产业经营组织、银行利益联结3方博弈模型,并分析3方合作过程中3阶段动态博弈过程,探寻有效降低金融支持农业产业发展不确定性带来的信用风险管理策略.     

Specific flavonoids as interconnecting signals in the tripartite symbiosis formed by arbuscular mycorrhizal fungi, Bradyrhizobium japonicum (Kirchner) Jordan and soybean (Glycine max (L.) Merr.)

Many legume plants benefit from the tripartite symbiosis of arbuscular mycorrhizal fungi (AMF) and rhizobia. Beneficial effects for the plant have been assumed to rely on increased P supply through the mycorrhizas. Recently, we demonstrated that P does not regulate the establishment of the tripartite symbiosis. Flavonoids appear to play a role as early signals for both rhizobia and AMF. Four soybean lines known to express different concentrations of the isoflavones genistein, daidzein and glycitein in the seed were used to test three hypotheses: (i) The establishment of the tripartite symbiosis is not dependent of a nutrient mediated effect; (ii) There is a positive correlation between seed isoflavone concentrations of different soybean lines and the progress of the tripartite symbiosis; (iii) Specific flavonoids control the establishment of the tripartite symbiosis in that a change in flavonoid root accumulation resulting from the development of one microbial partner can stimulate colonization of soybean roots by the other. Disturbed versus undisturbed soil treatments were produced to vary the potential for indigenous AMF colonization of soybean. In contrast, the potential for Bradyrhizobium was kept identical in both soil disturbance treatments. The uptake of P and Zn and the concentration of flavonoids in mycorrhizal soybean roots at 10 d after emergence were analysed either separately of Bradyrhizobium or in context of the tripartite symbiosis. Zinc nutrition did not differ between AMF treatments which supports the first hypothesis. The concentration of daidzein was at least four times greater in the root than in the seed reaching 3958±249 μg g⁻¹ dry across soybean lines. Coumestrol, which was absent in the seed, was synthesized to reach 2154±64 μg g⁻¹ dry. Conversely, the concentration of genistein was approximately three times smaller in the root that in the seed (301±15 μg g⁻¹ dry), while glycitein and formononetin were never detected. The establishment of the tripartite symbiosis was identical across soybean lines which does not support the second hypothesis. Concentrations of flavonoids were significantly greater in roots under disturbed soil, for which both symbioses were not as developed as in plants from undisturbed soil. This clearly supports the third hypothesis. This research provides the first data linking the function of different flavonoids to the establishment of the tripartite symbiosis, and suggests that these compounds are produced and released into the rhizosphere as a function of the colonization process.     

Role of TRIM8 in mouse cardiomyocyte apoptosis induced by high glucose and high free fatty acid

AIM: To investigate the effects of tripartite motif-containing protein 8 (TRIM8) on the apoptosis of mouse cardiomyocytes (MCMs) induced by high glucose and high free fatty acid (HGHF) and the underlying mechanism. METHODS: The MCMs were divided into normal glucose (NG) group (glucose at 5.5 mmol/L), high glucose (HG) group (glucose at 33 mmol/L), high free fatty acid (HF) group (sodium palmitate at 300 μmol/L) and HGHF group (glucose at 33 mmol/L and sodium palmitate at 300 μmol/L). The expression of TRIM8 in the MCMs was knocked down by siRNA, and the MCMs was further divided into control group, scrambled siRNA (Scra-siRNA)/PBS group, TRIM8-siRNA/PBS group, Scra-siRNA/HGHF group and TRIM8-siRNA/HGHF group. To further confirm the specific mechanism of TRIM8 in the MCM injury induced by HGHF, the MCMs were subgrouped into HGHF/DMSO group, HGHF+TRIM8-siRNA+DMSO (HGHF+Ts/DMSO) group, HGHF/ML385 group and HGHF+Ts/ML385 group. Accordingly, apoptosis was analyzed by flow cytometry, and the levels of reactive oxygen species (ROS) were measured by flow cytometry and DHE staining. The expression of TRIM8, nuclear factor E2-related factor 2 (Nrf2), glutamate-cysteine ligase catalytic subunit (GCLC), heme oxygenase-1 (HO-1) and NAD(P)H:quinone oxidoreductase 1 (NQO-1) at mRNA and protein levels was determined by qPCR and Western blot. RESULTS: HGHF increased the expression of TRIM8, and suppressed the expression of Nrf2, GCLC, HO-1 and NQO-1 in the MCMs (P < 0.05). Compared with Scra-siRNA/HGHF group, the intracellular ROS content and apoptotic rate were decreased in TRIM8-siRNA/HGHF group (P < 0.05). Correspondingly, the expression of the antioxidant molecule Nrf2 and its downstream genes GCLC, HO-1 and NQO-1 was increased (P < 0.05). In contrast, the addition of Nrf2 inhibitor ML385 partially reversed the inhibitory effect of TRIM8 expression knock-down on HGHF-induced apoptosis of MCMs. CONCLUSION: TRIM8 exacerbates the HGHF-induced cardiomyocyte apoptosis by modulating Nrf2 antioxidative pathway.     

The role of arbuscular mycorrhizal colonization in the carbon and nutrient economy of the tripartite symbiosis with nodulated Phaseolus vulgaris

In the tripartite symbiosis between nodulated legume roots and arbuscular mycorrhizal (AM) fungi, symbiont sink strength may depend upon developmental stage and the nutrient benefits to the host plant. The cost-benefits of the tripartite symbiosis were investigated in terms of C-economy and nutrition. Nodulated Phaseolus vulgaris seedlings, with and without AM, were hydroponically grown under high (2 mM) and low (1 μM) P conditions in an N-free Long Ashton nutrient solution. Plants were sequentially harvested at 17, 24 and 31 days after emergence. At each harvest, measurements for biomass, N₂-fixation, photosynthesis, root respiration, calculated C and nutritional economy were taken. Nodular growth was suppressed by the early development of AM colonization. This coincided with higher photosynthetic and respiratory rates in AM plants. These effects were most pronounced under low P when AM colonization peaked. Once AM levels reached the plateau phase, the efficiency of P nutrition increased. This was followed by improved nodular and host growth and enhanced N₂-fixation. This indicates that the AM was the dominant symbiont for host C in the tripartite symbiosis, due to its rapid development and subsequent role in supplying P more effectively to both host and nodules.     

Effect of TRIM44 silencing on proliferation of hepatocellular carcinoma cells and its molecular mechanism

ATM: To investigate the effect of tripartite motif-containing protein 44 (TRIM44) on the proliferation of hepatocellular carcinoma (HCC) cells and to study the molecular mechanism. METHODS: The expression of TRIM44 at mRNA and protein levels in normal liver tissues, HCC tissues, adjacent nontumor liver tissues, immortalized hepatocytes and hepatoma cell lines was determined by RT-qPCR and Western blot, respectively. The silencing of TRIM44 was conducted by transfection of vector expressing shRNA targeting TRIM44 (shTRIM44) in the HCC cells, and the protein level of TRIM44 was measured by Western blot. The viability of the HCC cells was analyzed by MTS assay. The DNA synthesis of HCC cells was detected by Click-iT EdU Imaging Kit. The ability of anchorage-independent growth was determined by the method of colony formation on the soft agar. The effects of TRIM44 on the total protein and phosphorylation of mammalian target of rapamycin (mTOR) levels were measured by Western blot. The HCC cells were transfected with shTRIM44 and treated with mTOR agonist MHY1485, and the cell viability was analyzed by MTS assay. RESULTS: The mRNA and protein levels of TRIM44 in the HCC tissues were significantly higher than those in the adjacent nontumor liver tissues and normal liver tissues. In addition, the mRNA and protein levels of TRIM44 in the hepatoma cell lines were significantly higher than those in the immortalized hepatocytes. TRIM44 silencing significantly inhibited the viability of HCC cells and reduced the abilities of DNA synthesis and anchorage-independent growth of the HCC cells. TRIM44 silencing decreased the phosphorylation level of mTOR protein. MHY1485 significantly antagonized the inhibitory effect of TRIM44 silence to the viability of HCC cells. CONCLUSION: TRIM44 silencing inhibits the proliferation of HCC cells possibly through down-regulating the activity of mTOR.     

TRIM25 ubiquitinates IGF2BP3 and inhibits viability of esophageal carcinoma EC109 cells

AIM: To investigate the relationship among tripartite motif-containing protein 25 (TRIM25), insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) and viability of human esophageal carcinoma EC109 cells. METHODS: The EC109 cells were divided into TRIM25 over-expression group, IGF2BP3 over-expression group, TRIM25 knock-down group, and IGF2BP3 knock-down and TRIM25 over-expression group. The viability of EC109 cells was mea-sured by MTT assay and CCK-8 assay. The stability of IGF2BP3 was detected by Western blot. The interaction between TRIM25 and IGF2BP3 was evaluated by immunoprecipitation, and the TRIM25 vector or empty vector was transfected to detect the ubiquitination of IGF2BP3 by immunoprecipitation. RESULTS: Over-expression of TRIM25 inhibited, but over-expression of IGF2BP3 promoted the viability of EC109 cells. However, the viability of the cells with knock-down of IGF2BP3 and over-expression of TRIM25 was lower than that of the cells with knock-down of IGF2BP3 only. Over-expression of TRIM25 resulted in reducing the expression level of IGF2BP3, which was recovered if the cells were treated with MG132, a proteasome inhibitor. When TRIM25 expression was knocked down, the viability of EC109 cells was significantly promoted on days 2, 3 and 4. The interaction between TRIM25 and IGF2BP3 was confirmed. At the same time, over-expression of TRIM25 increased the ubiquitination degree of IGF2BP3 in the EC109 cells. CONCLUSION: TRIM25 ubi-quitinates IGF2BP3, resulting in degradation of IGF2BP3 by proteasomes, thereby inhibiting the viability of esophageal carcinoma cells.