Yonghong Zhou

Negative regulation of CCaMK is essential for symbiotic infection

One of the earliest responses of legumes to symbiotic signalling is oscillation of the calcium concentration in the nucleoplasm of root epidermal cells. Integration and decoding of the calcium-spiking signal involve a calcium- and calmodulin-dependent protein kinase (CCaMK) and its phosphorylation substrates, such as CYCLOPS. Here we describe the Lotus japonicus ccamk-14 mutant that originated from a har1-1 suppressor screen. The ccamk-14 mutation causes a serine to asparagine substitution at position 337 located within the calmodulin binding site, which we determined to be an in vitro phosphorylation site in CCaMK. We show that ccamk-14 exerts cell-specific effects on symbiosis. The mutant is characterized by an increased frequency of epidermal infections and significantly compromised cortical infections by Mesorhizobium loti and also the arbuscular mycorrhiza fungus Rhizophagus irregularis. The S337 residue is conserved across angiosperm CCaMKs, and testing discrete substitutions at this site showed that it participates in a negative regulation of CCaMK activity, which is required for the cell-type-specific integration of symbiotic signalling.

Optimization of extraction process and antioxidant activity of polysaccharides from leaves of Paris polyphylla

Based on a single-factor test, a central composite design was used to optimize the extraction conditions of polysaccharides from leaves of Paris polyphylla Smith var. yunnanensis (Franch.) Hand.-Mazz. Three independent variables, including extraction temperature (°C), ratio of water to raw material, and extraction time (h), which significantly affected the yield of polysaccharides, were investigated. The experimental data were fitted to a quadratic polynomial equation using multiple regression analysis and also examined using appropriate statistical methods. The optimum conditions were as follows: extraction temperature, 90.8°C; ratio of water to raw material, 21.3:1; and extraction time 4.8h. Under these conditions, the experimental yield was 54.18%, which matched the predicted value well. Furthermore, the purified polysaccharide exerted strong antioxidant effects on DPPH, hydroxyl, and superoxide radicals in vitro.

Characterization, antioxidant and antitumor activities of polysaccharides from Salvia miltiorrhiza Bunge.

Polysaccharides were extracted from Salvia miltiorrhiza Bunge using response surface methodology (RSM) with ultrasonication. A Box-Behnken design was used to optimize the extraction parameters to maximize the polysaccharide extraction yield. The polysaccharide SMP-U1 was isolated and characterized; then the antioxidant and antiproliferation activities were evaluated in vitro. The modified optimal conditions were an ultrasonic power of 180 W, an extraction temperature of 54°C, and an extraction time of 32 min, achieving an extraction yield of 40.54±0.25%. The results indicate that SMP-U1 has significant antioxidant activity, scavenging the free radical 2,2-diphenyl-1-picrylhydrazyl. It has also exhibited effect on the proliferation of human breast carcinoma cells Bcap-37 and human esophageal carcinoma cells Eca-109, especially at a concentration of 0.30 mg/mL. In conclusion, SMP-U1 has remarkable in vitro antioxidant and antiproliferation activity, and has potential for application as a natural antioxidant or antitumor agent.

Ultrasonic-assisted extraction and antioxidant activities of polysaccharides from Camellia oleifera leaves.

Polysaccharides from Camellia oleifera leaves were extracted by ultrasonic-assisted extraction and optimized with response surface methodology. The optimal conditions of the extraction of polysaccharides were determined to be the extraction time of 1.6h, extraction temperature of 88°C and ratio of liquid to raw material of 20, which contributed to the highest yield of 3.77%. The crude polysaccharide was purified and three fractions (CLP-1, CLP-2 and CLP-3) were obtained. Their physicochemical properties were investigated with chemical method. The results indicated that the uronic acid content of CLP-2 was significantly higher than those of CLP-1 and CLP-3, and CLPs were mainly composed of glucose and galactose residue. In vitro antioxidant assay indicated that three polysaccharide fractions showed remarkable antioxidant activities in a concentration-dependent manner. Compared with CLP-1 and CLP-2, CLP-3 exhibited higher iron chelating activity. CLP-2 possessed significant higher hydroxyl radial and 2,2-diphenyl-1-picrydrazy (DPPH) radical scavenging activities than those of CLP-1 and CLP-3. Overall, the polysaccharides derived from C. oleifera leaves might be explored as potential natural antioxidant.

Optimization of extraction and antioxidant activity of polysaccharides from Salvia miltiorrhiza Bunge residue.

In this study, the process of extracting polysaccharides from Salvia miltiorrhiza Bunge residue was optimized by using a Box-Behnken design. Statistical analysis of the results showed that the linear and quadratic terms of the three variables of the extraction process had significant effects. The optimal conditions are as follows: extracting time of 2.6 h, extraction temperature of 89 °C, and ratio of water to raw material of 32 mL/g. Moreover, a new polysaccharide with antioxidant activity [i.e., SMWP-1 (∼5.27×10(5) Da)] was isolated from S. miltiorrhiza residue. The carbohydrate, uronic acid, and protein contents of SMWP-1 were 90.11%, 0.13%, and 0.53%, respectively. The SMWP-1 is composed of glucose, xylose, mannose, and galactose. The preliminary structural characterization of SMWP-1 was determined via Fourier transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM) analyses. This polysaccharide exhibited strong reducing power and free-radical scavenging activities in vitro against 2,2-diphenyl-1-picrylhydrazyl, superoxide anion, and hydroxyl. Therefore, SMWP-1 can be investigated further as a novel natural antioxidant.

Effect of exogenous salicylic acid on manganese toxicity, mineral nutrients translocation and antioxidative system in polish wheat (Triticum polonicum L.)

The present study investigated the morphological and physiological effect of salicylic acid (SA) on manganese toxicity in dwarf polish wheat (Triticum polonicum L.) seedlings grown hydroponically. Our findings showed that Mn stress could decrease plant growth, cause serious chlorosis and injury the photosynthetic apparatus. An increase of Mn accumulation and the inhibition of the K and Ca absorption and the Mg, Fe and Zn translocation were observed under Mn stress. Also, there was a considerable increase in H2O2 and TBARS (thiobarbituric acid-reactive substances) content in both the roots and leaves under Mn condition. The combination of SA and Mn treatment decreased the transport of Mn, Fe and Zn from roots to shoots and increased the Ca absorption and Mg translocation. In antioxidant system, such as CAT, APX, GR, DHAR, GSH and AsA, the combined treatment significantly increased the antioxidant content and antioxidative enzyme activities compared to the Mn stress alone. The level of ROS and lipid peroxidation significantly decreased under the combination of SA and Mn. These results suggested that SA-induced Mn tolerances in polish wheat are mainly by inhibiting Mn translocation, enhancing enzymatic activities and nonenzymatic antioxidants contents, and regulating nutrient absorption and distribution in plants.

Evolutionary pattern of rDNA following polyploidy in Leymus (Triticeae: Poaceae).

Ribosomal ITS polymorphism and its ancestral genome origin of polyploid Leymus were examined to infer the evolutionary outcome of rDNA gene following allopolyploid speciation and to elucidate the geographic pattern of ITS variation. The results demonstrated that different polyploids have experienced varying fates, including maintenance or homogenization of divergent arrays, occurrence of chimeric repeats and potential pseudogenes. Our data suggested that (1) the Ns, P/F, and St genomic types in Leymus were originated from Psathyrostachys, Agropyron/Eremopyrum, and Pseudoroegneria, respectively; (2) the occurrence of a higher proportion of Leymus species with predominant uniparental rDNA type might associate with the segmental allopolyploid origin, nucleolar dominance of alloploids, and rapid radiation of Leymus; (3) maintenance of multiple parental ITS types in allopolyploid might result from long generation times associated to vegetative multiplication, number and chromosomal location of ribosomal loci and/or recurrent hybridization; (4) the rDNA genealogical structure of Leymus species might associate with the geographic origins; and (5) ITS sequence clade shared by Leymus species from Central Asia, North America, and Nordic might be an outcome of ancestral ITS homogenization. Our results shed new light on understanding evolutionary outcomes of rDNA following allopolyploid speciation and geographic isolation.

Phylogenetic relationships in Leymus (Triticeae; Poaceae): Evidence from chloroplast trnH-psbA and mitochondrial coxII intron sequences

Leymus Hochst. is a polyploid genus with a diverse array of morphology, cytology, ecology, and distribution in Triticeae. To investigate the phylogenetic relationships and maternal genome donor of polyploid Leymus, the chloroplast trnH‐psbA region and mitochondrial coxII intron sequences of 33 Leymus taxa were analyzed with those of 36 diploid perennial species representing 19 basic genomes in Triticeae. The results showed that reticulate evolution occurred in Leymus species, with the cytoplasmic lineage of Leymus contributed by different progenitors. Interspecific relationships of Leymus were also elucidated on the basis of orthologous comparison. Our data suggested that: (i) due to incomplete lineage sorting and/or difference in the pattern of chloroplast and mitochondrial inheritance, the genealogical conflict between the two genealogical patterns suggest the contribution of Psathyrostachys Nevski, Agropyron J. Gaertn, Eremopyrum (Ledeb.) Jaub. & Spach, Pseudoroegneria (Nevski) Á. Löve, Thinopyrum Á. Löve, and Lophopyrum (Host) Á. Löve to the cytoplasmic lineage of Leymus; (ii) there is a close relationship among Leymus species from the same area or neighboring geographic regions; (iii) L. coreanus (Honda) K. B. Jensen & R. R.‐C. Wang, L. duthiei (Stapf) Y. H. Zhou & H. Q. Zhang ex C. Yen, J. L. Yang & B. R. Baum, L. duthiei var. longearistatus (Hack.) Y. H. Zhou & H. Q. Zhang ex C. Yen, J. L. Yang & B. R. Baum, and L. komarovii (Roshev.) C. Yen, J. L. Yang & B. R. Baum are closely related to other Leymus species, and it is reasonable to transfer these species from the genus Hystrix Moench to Leymus; (iv) Leymus species from North America are closely related to L. coreanus from the Russian Far East and L. komarovii from northeast China but are evolutionarily distinct from Leymus species from Central Asia and the Qinghai–Tibet Plateau. The occurrence of multiple origin and introgression could account for the rich diversity and ecological adaptation of Leymus species.

Phylogeny and differentiation of the St genome in Elymus L. sensu lato (Triticeae; Poaceae) based on one nuclear DNA and two chloroplast genes.

BackgroundHybridization and polyploidization can be major mechanisms for plant evolution and speciation. Thus, the process of polyploidization and evolutionary history of polyploids is of widespread interest. The species in Elymus L. sensu lato are allopolyploids that share a common St genome from Pseudoroegneria in different combinations with H, Y, P, and W genomes. But how the St genome evolved in the Elymus s. l. during the hybridization and polyploidization events remains unclear. We used nuclear and chloroplast DNA-based phylogenetic analyses to shed some light on this process.ResultsThe Maximum likelihood (ML) tree based on nuclear ribosomal internal transcribed spacer region (nrITS) data showed that the Pseudoroegneria, Hordeum and Agropyron species served as the St, H and P genome diploid ancestors, respectively, for the Elymus s. l. polyploids. The ML tree for the chloroplast genes (matK and the intergenic region of trnH-psbA) suggests that the Pseudoroegneria served as the maternal donor of the St genome for Elymus s. l. Furthermore, it suggested that Pseudoroegneria species from Central Asia and Europe were more ancient than those from North America. The molecular evolution in the St genome appeared to be non-random following the polyploidy event with a departure from the equilibrium neutral model due to a genetic bottleneck caused by recent polyploidization.ConclusionOur results suggest the ancient common maternal ancestral genome in Elymus s. l. is the St genome from Pseudoroegneria. The evolutionary differentiation of the St genome in Elymus s. l. after rise of this group may have multiple causes, including hybridization and polyploidization. They also suggest that E. tangutorum should be treated as C. dahurica var. tangutorum, and E. breviaristatus should be transferred into Campeiostachys. We hypothesized that the Elymus s. l. species origined in Central Asia and Europe, then spread to North America. Further study of intraspecific variation may help us evaluate our phylogenetic results in greater detail and with more certainty.

Cadmium Treatment Alters the Expression of Five Genes at the Cda1 Locus in Two Soybean Cultivars [Glycine Max (L.) Merr]

Westag 97 has larger capacity of Cd accumulation in roots which prevents Cd from translocating into stems and leaves; conversely, AC Hime has smaller capacity of Cd accumulation in roots; more Cd is transported into stems and leaves. The different capacity of Cd in roots between Westag 97 and AC Hime causes the different Cd concentration in seeds. Meanwhile, according to the different expression levels of RSTK, ISCP, and H+-ATPase between Westag 97 and AC Hime, RSTK may be involved in transporting Cd into stems and leaves; H+-ATPase may be correlated to the capacity of Cd accumulation in roots; and Cd caused some changes of fundamental life process which leaded to the different expression patterns of ISCP between Westag 97 and AC Hime.