Disruption of LLM9428/OsCATC Represses Starch Metabolism and Confers Enhanced Blast Resistance in Rice.

Catalases (CATs) are important self-originating enzymes and are involved in many of the biological functions of plants. Multiple forms of CATs suggest their versatile role in lesion mimic mutants (LMMs), H2O2 homeostasis and abiotic and biotic stress tolerance. In the current study, we identified a large lesion mimic mutant9428 (llm9428) from Ethyl-methane-sulfonate (EMS) mutagenized population. The llm9428 showed a typical phenotype of LMMs including decreased agronomic yield traits. The histochemical assays showed decreased cell viability and increased reactive oxygen species (ROS) in the leaves of llm9428 compared to its wild type (WT). The llm9428 showed enhanced blast disease resistance and increased relative expression of pathogenesis-related (PR) genes. Studies of the sub-cellular structure of the leaf and quantification of starch contents revealed a significant decrease in starch granule formation in llm9428. Genetic analysis revealed a single nucleotide change (C > T) that altered an amino acid (Ala > Val) in the candidate gene (Os03g0131200) encoding a CATALASE C in llm9428. CRISPR-Cas9 targetted knockout lines of LLM9428/OsCATC showed the phenotype of LMMs and reduced starch metabolism. Taken together, the current study results revealed a novel role of OsCATC in starch metabolism in addition to validating previously studied functions of CATs.

Rapid Visual Detection of High Nitrogen-Use Efficiency Gene OsGRF4 in Rice (Oryza sativa L.) Using Loop-Mediated Isothermal Amplification Method.

The GROWTH-REGULATING FACTOR4 (OsGRF4) allele is an important target for the development of new high nitrogen-use efficiency (NUE) rice lines that would require less fertilizers. Detection of OsGRF4 through PCR (polymerase chain reaction)-based assay is cumbersome and needs advanced laboratory skills and facilities. Hence, a method for conveniently and rapidly detecting OsGRF4 on-field is a key requirement for further research and applications. In this study, we employed cleaved amplified polymorphic sequences (CAPs) and loop-mediated isothermal amplification (LAMP) techniques to develop a convenient visual detection method for high NUE gene OsGRF4NM73 (OsGRF4 from the rice line NM73). The TC→AA mutation at 1187-1188 bp loci was selected as the target sequence for the OsGRF4NM73 allele. We further employed this method of identification in 10 rice varieties that carried the OsGRF4 gene and results revealed that one variety (NM73) carries the target OsGRF4NM73 allele, while other varieties did not possess the osgrf4 genotype. The optimal LAMP reaction using hydroxynaphthol blue (HNB), a chromogenic indicator, was carried out at 65 °C for 60 min, and the presence of OsGRF4NM73 allele was confirmed by color changes from violet to sky blue. The results of this study showed that the LAMP method can be conveniently and accurately used to detect the OsGRF4NM73 gene in rice.

Quantifying the effects of plant density on soybean lodging resistance and growth dynamics in maize-soybean strip intercropping.

Shading-induced soybean stem lodging is a prevalent concern in the maize (Zea mays L.)-soybean (Glycine max L. Merr.) strip intercropping system, leading to a substantial decline in yield. Nevertheless, the associations between soybean growth, stem lodging, and yield formation in this scenario remain unclear. To investigate this, the logistic and beta growth models were utilized to analyze the growth process of soybean organs (stems, leaves, branches, and pods) and the accumulation of carbohydrates (lignin, cellulose, and sucrose) at three planting densities (8.5, 10, and 12.5 plants m-2) in both strip intercropping and skip strip monoculture systems. The results indicate that shading stress caused by maize in the intercropping system reduced lignin and cellulose accumulation in soybean stems, thus decelerating soybean organ growth compared to monoculture. Furthermore, intercropped soybean at higher planting density (PD3) exhibited a 28% reduction in the maximum dry matter growth rate (cm) and a 11% decrease in the time taken to reach the maximum dry matter growth rate (te) compared to the lower planting density (PD1). Additionally, a 29% decrease in the maximum accumulation rate (cmax) of sucrose, lignin, and cellulose was observed, along with a 13% decrease in the continuous accumulation time (tc) of these carbohydrates in intercropped soybean at PD3. Interspecific and intraspecific shading stress led to a preferential allocation of assimilates into soybean stems, enhancing plant height during the initial stage, while at later stages, a greater proportion of sucrose was allocated to leaves. Consequently, this hindered the conversion of sucrose into lignin and cellulose within the stems, ultimately resulting in a reduction in the lodging resistance index (LRI). Overall, this study provides valuable insights into the effects of shading stress on soybean growth and yield. It also emphasizes how optimizing planting density in intercropping systems can effectively alleviate shading stress and enhance crop productivity.

Recombinant Viral Capsid Protein L2 (rVL2) of HPV 16 Suppresses Cell Proliferation and Glucose Metabolism via ITGB7/C/EBPß Signaling Pathway in Cervical Cancer Cell Lines.

PURPOSE: Capsid protein L2 is the minor capsid protein of human papillomavirus 16 (HPV16). Although L2-based vaccines were developed, the therapeutic effect of recombinant viral capsid protein L2 (rVL2) was still to be illustrated. METHODS: We used glucose uptake and lactate production assay to verify the inhibitory effect of rVL2 on the glucose metabolism in cervical cancer cells. Secondly, we performed gene-chip assay, RT-PCR, and Western blot to determine the role of ITGB7/C/EBPß signaling pathway in rVL2-mediated glucose metabolism in vitro. Finally, we used an animal model to verify the function of rVL2 in cervical cancer. RESULTS: We found that rVL2 reduced glucose uptake and lactate production levels in cervical cancer cells, which caused the inhibition of cell proliferation. rVL2 decreased the expression levels of key metabolic enzymes, including GLUT1, LDHA, and ALDOA, to affect cell metabolism in cervical cancer cells by inhibiting ITGB7/C/EBPß signaling pathway in vitro and in vivo. CONCLUSION: These results demonstrated the vital role of rVL2 in the glycolysis-induced cell growth and proliferation via suppressing ITGB7/C/EBPß signaling axis.

MiR-449a functions as a tumor suppressor in endometrial cancer by targeting CDC25A.

Accumulating evidence has demonstrated that microRNAs (miRNAs) play critical roles in cancer initiation and development by functioning either as oncogenes or as tumor suppressors. The role of microRNA-449a (miR-449a) in endometrial cancer remains unclear. We examined the levels of miR-449a and miR-449b in benign endometrium, type I and type II endometrial cancer tissues by quantitative real-time polymerase chain reaction. To further investigate the roles of miR-449a in regulating the behavior of endometrial cancer cells, we overexpressed miR-449a in the endometrial cancer cell line HEC-1B, which had low endogenous miR-449a expression. We analyzed the effects of miR-449a overexpression on CDC25 expression, proliferation, invasion and apoptosis of HEC-1B cells. We found that miR-449a and miR-449b levels were markedly reduced in type II endometrial cancer tissues but not in type I endometrial cancer tissues compared with normal endometrium. Overexpression of miR-449a significantly inhibited the proliferation, invasion and clonogenic survival of HEC-1B cells. MiR-449a overexpression also induced apoptosis in HEC-1B cells. In addition, real-time RT-PCR and western blot analysis showed that CDC25A expression was suppressed by miR-449a overexpression. Our results suggest that miR-449a may act as a tumor suppressor by targeting CDC25A in endometrial cancer.

[Influence of penetrative needling of Shendao (GV 11) on the symptom score and serum IgE content in chronic urticaria patients].

OBJECTIVE: To observe the changes of symptom scores and serum IgE level after treatment with thick-needle subcutaneous penetration of Shendao (GV 11) in chronic urticaria patients. METHODS: A total of 60 chronic urticaria patients were randomly divided into acupuncture group (n = 30) and medication group (n = 30). Subcutaneous penetrative needling was applied to GV 11 with thick acupuncture needle (retained for 4 h/time, once daily, 5 times/week) for patients of acupuncture group and Levocetirizine Hydrochloride tablets (5 mg/time, once daily in the first two weeks, then, once every other day in the 3rd and 41th weeks, and once every 3 days in the last two weeks) were given to patients of medication group. Serum IgE content was assayed before and 2,6, 12 weeks after the treatment by chemiluminescent technique. Symptom scores were obtained by "0-3 four levels assessment" method in the light of the size of the wheal and the itching severity. RESULTS: Self-comparison indicated that the symptom scores and serum IgE levels declined significantly (P < 0.01) 2 and 6 weeks (Wks) after the treatment in both acupuncture and medication groups,and 12 Wks after the treatment in the acupuncture group (P < 0.01). Comparison between two groups showed that the symptom score and serum IgE content of acupuncture group were significant lower than those of medication group 12 Wks after the treatment (P < 0.05). No significant differences were found between two groups in the symptom scores and serum IgE levels before, 2 and 6 Wks after the treatment (P > 0.05). A positive correlation exists between the symptom score and the serum IgE level before and after the treatment in both groups. CONCLUSION: Thick-needle subcutaneous penetration of Shendao (GV 11) can effectively improve clinical symptoms of chronic urticaria patients, which may be closely related to its effect in lowering peripheral blood IgE level.