Characterization of five CIPK genes expressions in maize under water stress

ABA, H₂O₂ and Ca²⁺ play critical roles as signals in the adaptive responses of plants to water and other stresses. They accumulate in plant cells under water and other stresses and induce changes in stress-related gene expressions. CIPKs, protein kinases associated with a calcineurin B-like calcium sensor, play a role in the regulation of stress gene expression in plants. However, it is still unclear whether ABA and H₂O₂ are key inducers that regulate the changes in CIPK expressions under water stress. In this study, five stress-inducible CIPKs in maize were retrieved from Database. They were designated as ZmCIPK1, 3, 8, 17 and 18, based on their homologies with known CIPK sequences. The expressions of the five ZmCIPKs in maize leaves and roots were analyzed and found to be regulated by PEG, CaCl₂, ABA and H₂O₂ to different extents. Moreover, the effect of ABA and H₂O₂ on the expressions of ZmCIPKs under water stress was in an organ-dependent manner.     

A mutation in a coproporphyrinogen III oxidase gene confers growth inhibition, enhanced powdery mildew resistance and powdery mildew-induced cell death in Arabidopsis

Key message

A gene encoding a coproporphyrinogen III oxidase mediates disease resistance in plants by the salicylic acid pathway.

Abstract

A number of genes that regulate powdery mildew resistance have been identified in Arabidopsis, such as ENHANCED DISEASE RESISTANCE 1 to 3 (EDR1 to 3). To further study the molecular interactions between the powdery mildew pathogen and Arabidopsis, we isolated and characterized a mutant that exhibited enhanced resistance to powdery mildew. The mutant also showed dramatic powdery mildew-induced cell death as well as growth defects and early senescence in the absence of pathogens. We identified the affected gene by map-based cloning and found that the gene encodes a coproporphyrinogen III oxidase, a key enzyme in the tetrapyrrole biosynthesis pathway, previously known as LESION INITIATION 2 (LIN2). Therefore, we designated the mutant lin2-2. Further studies revealed that the lin2-2 mutant also displayed enhanced resistance to Hyaloperonospora arabidopsidis (H.a.) Noco2. Genetic analysis showed that the lin2-2-mediated disease resistance and spontaneous cell death were dependent on PHYTOALEXIN DEFICIENT 4 (PAD4), SALICYLIC ACID INDUCTION-DEFICIENT 2 (SID2), and NONEXPRESSOR OF PATHOGENESIS-RELATED GENES 1 (NPR1), which are all involved in salicylic acid signaling. Furthermore, the relative expression levels of defense-related genes were induced after powdery mildew infection in the lin2-2 mutant. These data indicated that LIN2 plays an important role in cell death control and defense responses in plants.     

Proposal of Ensifer psoraleae sp. nov., Ensifer sesbaniae sp. nov., Ensifer morelense comb. nov. and Ensifer americanum comb. nov

In a survey of rhizobia associated with the native legumes in Yunnan Province, China, seven and nine strains isolated from the root nodules of Psoralea corylifolia, Sesbania cannabina and Medicago lupulina were respectively classified into the novel genomic species groups I and II in the genus Ensifer (former Sinorhizobium) based on the sequence analyses of the 16S rRNA gene. Analyses of concatenated housekeeping genes (atpD, recA and glnII) further revealed that they were distinct lineages in the genus, and group I was most similar to Ensifer terangae and Ensifer garamanticus (both with 94.2% similarity), while group II was most similar to Ensifer adhaerens (94.0%). These groups could be distinguished from closely related species by DNA–DNA relatedness, MALID-TOF MS, cellular fatty acid profiles and a series of phenotypic characters. Therefore, two novel species were proposed: Ensifer psoraleae sp. nov. (seven strains, type strain CCBAU 65732^T = LMG 26835^T = HAMBI 3286^T) and Ensifer sesbaniae sp. nov. (nine strains, type strain CCBAU 65729^T = LMG 26833^T = HAMBI 3287^T). They had a DNA G + C mol% (T_m) of 58.9 and 60.4, respectively. Both of the type strains formed effective nodules on common bean (Phaseolus vulgaris) and their hosts of origin. In addition, the previously described species Sinorhizobium morelense and Sinorhizobium americanum were renamed as Ensifer morelense comb. nov. and Ensifer americanum comb. nov. according to the accumulated data from different studies.     

Molecular Cloning and Expression Analysis of Nine ThTrx Genes in Tamarix hispida

Thioredoxins are small conserved proteins that play key roles in the oxidative stress response. In this study, nine Trx genes, including five Trxhs, three Trxms, and one Trx-like gene, were cloned from Tamarix hispida. The roles of these ThTrx genes were investigated under various abiotic stress conditions. Expression profiles of the nine ThTrx genes in response to different abiotic stresses in leaf and root tissues were constructed using quantitative real time-polymerase chain reaction. Differential expression of all nine ThTrx genes was observed (>2-fold) in response to NaCl, PEG, or CdCl₂ stress in at least one tissue, indicating that all of these genes act in abiotic stress responses. All ThTrx genes were induced (>2-fold) by abscisic acid (ABA) treatment in the leaves and especially in the roots, suggesting that ABA-dependent signaling pathways regulate ThTrxs. These results demonstrate that ThTrx expression constitutes an adaptive response to abiotic stress in T. hispida and plays an important role in abiotic stress tolerance.     

8 Using pseudorotation as a reaction coordinate in free energy simulations of nucleic acids

Backbone sugar groups are central components of nucleic acids. The conformations of the ribose/deoxyribose can be elegantly described using the concept of pseudorotation (Altona and Sundaralingam, 1972), and are dominated by the C2′- and C3′-endo conformers. The free energy barrier of the transition between these two major puckering modes can be probed by NMR relaxation experiments (Johnson and Hoogstraten, 2008), but an atomic picture of the transition path per se is only available for several truncated nucleoside analogues (Brameld & Goddard III, 1999). Here, we implemented a new free energy simulation method for Molecular Dynamics simulations using pseudorotation as the reaction coordinate (Cremer and Pople, 1975). This allowed us to compute the free energy landscape of a complete pseudorotation cycle. The free energy landscape revealed not only the relative stability of C2′- and C3′-endo conformers, but also the main transition path and its free energy barrier. As a validation of our new approach, we calculated free energy surface of the pseudorotation of guanosine monophosphate. The free energy surface revealed that the C2′-endo conformation is ?1?kcal/mol that is more stable and the free energy barrier for the transition is 4.5–5?kcal/mol. These are in excellent agreement with previous NMR measurements (Zhang et al., 2012; Röder et al., 1975). We have further applied this method to other systems that are important in pre-biotic chemistry, including an RNA duplex with unique 2′, 5′-phosphodiester linkages.     

Needle δ13C and mobile carbohydrates in Pinus koraiensis in relation to decreased temperature and increased moisture along an elevational gradient in NE China

A tree’s crown interacts with atmospheric variables such as CO₂, temperature, and humidity. Physioecology of leaves/needles (e.g. δ¹³C, mobile carbohydrates, and nitrogen) is, therefore, strongly affected by microclimate in and surrounding a tree crown. To understand the physiological responses of leaves to changes in air temperature and moisture, we measured δ¹³C, soluble sugars, starch, and total nitrogen (N) concentrations in current year and 1-yr-old needles of Pinus koraiensis trees, and compared the growing season air temperature and relative humidity within and outside P. koraiensis crowns along an elevational gradient from 760 to 1,420 m a.s.l. on Changbai Mountain, NE China. Our results indicated that needle N and mobile carbohydrates concentrations, as well as needle δ¹³C values changed continuously with increasing elevation, corresponding to a continuous decrease in air temperature and an increase in relative humidity. Needle carbon and nitrogen status is highly significantly negatively correlated with temperature, but positively correlated with relative humidity. These results indicate that increases in air temperature in combination with decreases in relative humidity may result in lower levels of N and mobile carbohydrates in P. koraiensis trees, suggesting that future climate changes such as global warming and changes in precipitation patterns will directly influence the N and carbon physiology at P. koraiensis individual level, and indirectly affect the competitive ability, species composition, productivity and functioning at the stand and ecosystem level in NE China. Due to the relatively limited range of the transect (760–1,420 m) studied, further research is needed to explain whether the present results are applicable to scales across large elevational gradients.