Arabidopsis CDPK6 phosphorylates ADF1 at N-terminal serine 6 predominantly

Key message

We found that Arabidopsis AtADF1 was phosphorylated by AtCDPK6 at serine 6 predominantly and the phosphoregulation plays a key role in the regulation of ADF1-mediated depolymerization of actin filaments.

Abstract

Since actin-depolymerizing factor (ADF) is highly conserved among eukaryotes, it is one of the key modulators for actin organization. In plants, ADF is directly involved in the depolymerization of actin filaments, and therefore important for F-actin-dependent cellular activities. The activity of ADF is tightly controlled through a number of molecular mechanisms, including phosphorylation-mediated inactivation of ADF. To investigate Arabidopsis ADF1 phosphoregulation, we generated AtADF1 phosphorylation site-specific mutants. Using transient expression and stable transgenic approaches, we analyzed the ADF1 phosphorylation mutants in the regulation of actin filament organizations in plant cells. By in vitro phosphorylation assay, we showed that AtADF1 is phosphorylated by AtCDPK6 at serine 6 predominantly. Chemically induced expression of AtCDPK6 can negatively regulate the wild-type AtADF1 in depolymerizing actin filaments, but not those of the mutants AtADF1(S6A) and AtADF1(S6D). These results demonstrate a regulatory function of Arabidopsis CDPK6 in the N-terminal phosphorylation of AtADF1.     

Molecular cloning and characterization of a Brassica juncea yellow stripe-like gene, BjYSL7, whose overexpression increases heavy metal tolerance of tobacco

Key message

BjYSL7 encodes a plasma-localized metal–NA transporter and has transport Fe(II)–NA complexes activity. BjYSL7 is involved in the transport of Cd and Ni from roots to shoots.

Abstract

Heavy metal transporters play a key role in regulating metal accumulation and transport in plants. In this study, we isolated a novel member of the yellow stripe-like (YSL) gene family BjYSL7 from the hyperaccumulator Brassica juncea. BjYSL7 is composed of 688 amino acids with 12 putative transmembrane domains and is over 90 % identical to TcYSL7 and AtYSL7. Real-time PCR analysis revealed that BjYSL7 mRNA was mainly expressed in the stem under normal condition. The expression of BjYSL7 was found to be up-regulated by 127.1-, 12.7-, and 3.4-fold in roots and 6.5-, 4.3-, and 2.8-fold in shoots under FeSO₄, NiCl₂, and CdCl₂ stresses, respectively. We have demonstrated that BjYSL7 is a Fe(II)–NA influx transporter by yeast functional complementation. Moreover, a BjYSL7::enhanced green fluorescent protein (EGFP) fusion localized to the plasma membrane of onion epidermal cells. The BjYSL7-overexpressing transgenic tobacco plants exhibited longer root lengths, lower relative inhibition rate of lengths and superior root hair development compared to that of wild-type (WT) plants in the presence of CdCl₂ and NiCl₂. Furthermore, the concentrations of Cd and Ni in shoots of BjYSL7-overexpressing plants are significantly higher than that of WT plants. Compared with WT plants, BjYSL7-overexpressing plants exhibited Fe concentrations that were higher in the shoots and seeds and lower in the roots. Taken together, these results suggest that BjYSL7 might be involved in the transport of Fe, Cd and Ni to the shoot and improving heavy metal resistance in plants.     

The effects of long-term fertilization on the accumulation of organic carbon in the deep soil profile of an oasis farmland

Background and aims

Deeper soils represent a poorly understood, but potentially important, sink for carbon sequestration. The objective of this study was to determine the effects of long-term fertilization on soil organic carbon (SOC), its labile fractions and aggregate-associated carbon throughout a 0–3 m soil profile.

Methods

The investigation was conducted in a field experiment started in 1990 in an oasis farmland cropped with winter wheat. The following treatments were compared with the desert from which the oasis was created: CK (no fertilizer), NPK, N2P2K, NPKR, and N2P2R2 (“2” for double fertilizer and “R” for straw residue)

Results

SOC contents increased by 14–56 % in the topsoil (0–0.2 m), but decreased by 15–22 % in the subsoil (0.2–0.6 m) under all fertilizer treatments. In the deep layer (0.6–3 m) there were significant differences between the treatments: SOC decreased by 5–9 % in treatments without straw, but increased by 4–9 % in treatments with straw. Labile fractions (particulate organic carbon and light fraction organic carbon) also showed similar trends. Both the fertilizer and CK treatments led to an increase in the amount of macro-aggregates (>0.25 mm), especially small macro-aggregates (0.25–2 mm), throughout the soil profile. SOC content was highest in the macro-aggregates, intermediate in the silt + clay fraction (<0.053 mm), and lowest in the micro-aggregates (0.25–0.053 mm). However, 44–87 % of total SOC was stored in the silt + clay fraction, especially in the deep layer (at least 80 %).

Conclusions

After 20 years of fertilizer applications, difference in SOC mainly occurred in the deep layer, and preservation of SOC in the silt + clay fraction appeared to be a prerequisite for soil-carbon sequestration. Applying inorganic fertilizer alone decreased SOC content in the silt + clay fraction in the deep layer, while the combined applications with straw resulted in higher SOC content in the silt + clay fraction in that layer, which turned out to be the main mechanism for increasing SOC content. Our study indicated that applying straw with inorganic fertilizer is the best practice for carbon sequestration, which occurred mainly in the deep soil layer.     

Summer rain pulses may stimulate a CO2 release rather than absorption in desert halophyte communities

Background and aims

The response of plants and soil to rain pulses determines seasonal variations in the exchange of materials and energy at the ecosystem scale in arid and semi-arid regions. We assessed how the ecosystem carbon exchange (NEE) of desert halophyte communities of different plant functional-types responds to summer precipitation pulses in Tamarix and Haloxylon communities.

Methods

Plant water status, photosynthetic gas exchange, soil respiration and net ecosystem carbon exchange were measured to test the hypothesis that high physiological sensitivity may induce a greater changes in NEE resulting from the summer precipitation pulses in Haloxylon community.

Results

Plant water status and photosynthetic assimilation did not differ before and after summer precipitation pulses in either community. In contrast, soil respiration and NEE responded strongly to summer precipitation events in both communities. At the ecosystem level, precipitation pulses induced a pulse of CO₂ release, rather than absorption. The NEE response to summer precipitation was less in the deep-rooted Tamarix community, compared to the shallow-rooted Haloxylon community, which was even converted into a carbon source after summer precipitation inputs. As a result, the effect of summer precipitation inputs on soil respiration was more important than the plant (carbon assimilation) response in determining the ecosystem response to episodic precipitation pulses.     

A core functional region of the RFP1 promoter from Chinese wild grapevine is activated by powdery mildew pathogen and heat stress

RING-finger proteins (RFP) function as ubiquitin ligases and play key roles in plant responses to biotic and abiotic stresses. However, little information is available on the regulation of RFP expression. Here, we isolate and characterize the RFP promoter sequence from the disease-resistant Chinese wild grape Vitis pseudoreticulata accession Baihe-35-1. Promoter-GUS fusion assays revealed that defense signaling molecules, powdery mildew infection, and heat stress induce VpRFP1 promoter activity. By contrast, the RFP1 promoter isolated from Vitis vinifera was only slightly induced by pathogen infection and heat treatment. By promoter deletion analysis, we found that the ?148 bp region of the VpRFP1 promoter was the core functional promoter region. We also found that, in Arabidopsis, VpRFP1 expressed under its own promoter activated defense-related gene expression and improved disease resistance, but the same construct using the VvRFP1 promoter slightly improve disease resistance. Our results demonstrated that the ?148 bp region of the VpRFP1 promoter plays a key role in response to pathogen and heat stress, and suggested that expression differences between VpRFP1 and VvRFP1 may be key for the differing disease resistance phenotypes of the two Vitis genotypes.     

Sodium Taurocholate Cotransporting Polypeptide Mediates Woolly Monkey Hepatitis B Virus Infection of Tupaia Hepatocytes

Primary Tupaia hepatocytes (PTHs) are susceptible to woolly monkey hepatitis B virus (WMHBV) infection, but the identity of the cellular receptor(s) mediating WMHBV infection of PTHs remains unclear. Recently, sodium taurocholate cotransporting polypeptide (NTCP) was identified as a functional receptor for human hepatitis B virus (HBV) infection of primary human and Tupaia hepatocytes. In this study, a synthetic pre-S1 peptide from WMHBV was found to bind specifically to cells expressing Tupaia NTCP (tsNTCP) and it efficiently blocked WMHBV entry into PTHs; silencing of tsNTCP in PTHs significantly inhibited WMHBV infection. Ectopic expression of tsNTCP rendered HepG2 cells susceptible to WMHBV infection. These data demonstrate that tsNTCP is a functional receptor for WMHBV infection of PTHs. The result also indicates that NTCP''s orthologs likely act as a common cellular receptor for all known primate hepadnaviruses.     

Infection Status of Hydatid Cysts in Humans and Sheep in Uzbekistan

Uzbekistan is endemic of cystic echinococcosis (CE). In order to estimate endemicity of CE, we collected data from emergency surgery due to CE in 2002-2010 and also investigated the prevalence of hydatid cysts in the liver and lungs of sheep at an abattoir in Uzbekistan from July 2009 to June 2010. In 14 emergency hospitals, 8,014 patients received surgical removal or drainage of CE during 2002-2010, and 2,966 patients were found in 2010. A total of 22,959 sheep were grossly examined of their liver and lungs, and 479 (2.1%) and 340 (1.5%) of them were positive for the cyst in the liver and lungs, respectively. Echinococcus granulosus is actively transmitted both to humans and sheep, and CE is a zoonotic disease of public health priority in Uzbekistan.