Jian-Hui Wang

Arabidopsis cryptochrome 1 functions in nitrogen regulation of flowering

Significance Overapplication of nitrogen (N) fertilizer causes delayed flowering and negatively impacts the function and composition of natural ecosystems and climate. In this study, we demonstrate that flowering time variations regulated by altered nitrogen levels are mediated by two key factors: ferredoxin-NADP+-oxidoreductase (FNR1) and the blue-light receptor cryptochrome 1 (CRY1). Nitrogen regulates FNR1 expression, thereby contributing to changes in NADPH/NADP+ and ATP/AMP ratios, which in turn activates adenosine monophosphate-activated protein kinase to modulate nuclear CRY1 abundance, which further acts in the N signal input pathway to affect central clock function and flowering time. A better understanding of N-regulated floral transition will offer biotechnological solutions to improve sustainable agriculture. The phenomenon of delayed flowering after the application of nitrogen (N) fertilizer has long been known in agriculture, but the detailed molecular basis for this phenomenon is largely unclear. Here we used a modified method of suppression-subtractive hybridization to identify two key factors involved in N-regulated flowering time control in Arabidopsis thaliana, namely ferredoxin-NADP+-oxidoreductase and the blue-light receptor cryptochrome 1 (CRY1). The expression of both genes is induced by low N levels, and their loss-of-function mutants are insensitive to altered N concentration. Low-N conditions increase both NADPH/NADP+ and ATP/AMP ratios, which in turn affect adenosine monophosphate-activated protein kinase (AMPK) activity. Moreover, our results show that the AMPK activity and nuclear localization are rhythmic and inversely correlated with nuclear CRY1 protein abundance. Low-N conditions increase but high-N conditions decrease the expression of several key components of the central oscillator (e.g., CCA1, LHY, and TOC1) and the flowering output genes (e.g., GI and CO). Taken together, our results suggest that N signaling functions as a modulator of nuclear CRY1 protein abundance, as well as the input signal for the central circadian clock to interfere with the normal flowering process.

The roles of two transcription factors, ABI4 and CBFA, in ABA and plastid signalling and stress responses

Genetic and physiological studies have revealed evidences for multiple signaling pathways by which the plastid exerts retrograde control over photosynthesis-associated-nuclear-genes. In this study we have examined the mechanisms of control of transcription by plastid signals, focusing on transcription factors. We have also further addressed the physical nature of plastid signals and the physiological role, in stress acclimation of this regulatory pathway. ABI4, a master Apetala 2 (AP2)-type transcription factor (TF), is targeted by multiple signalling pathways in plant cells, such as abscisic acid (ABA) signals, sugar signals and plastid signals derived from reactive oxygen species (ROS) and chlorophyll intermediates. ABI4 binds the promoter of target genes to prevent their transcription by competing with other competitive TFs. However, we found that once ABI4 bound the element (CCACGT), it may not be bound by other TFs, therefore making the signalling long-lasting. Downstream of ABI4, CBFA (CCAAT binding factor A) is a subunit of the HAP2/HAP3/HAP5 (Heme activator protein) trimeric transcription complex. CBFA however is a redundant HAP3 subunit. When emergency occurs (such as herbicide treatments or environmental stresses followed by ABA and ROS accumulation), the master transcription factor ABI4 down-regulates some TFs, like CBFA, and then some other TF subunits enter the transcription complex and transcriptional efficiency of stress-responsive genes (including the transcription co-factor CBP) is improved instantaneously. abi4, cbfA and cbp mutants showed weaker drought-tolerance after a herbicide norflurazon treatment, which indicated the physiological role of these key transcription factors.

Variation analysis of two cucumber mosaic viruses and their associated satellite RNAs from sugar beet in China

Two cucumber mosaic virus (CMV) isolates XJ1 and XJ2 were obtained from sugar beet showing yellow mosaic symptom in Shihezi, Xinjiang Uigur municipality of China. The coat protein gene of the two CMV isolates and their associated satellite RNAs were amplified by reverse transcriptase polymerase chain reaction (RT-PCR) and were cloned and sequenced. Comparison of CP gene sequences showed that XJ1 and XJ2 have the highest sequence identity with that of CMV-Danshen (97.8%) and CMV-SD (98.7%), respectively. Two types of satellite RNAs (XJs1 and XJs2) were found to be associated with the two CMV isolates consisting of 384 nucleotides and 336 nucleotides, respectively. Sequence comparisons revealed that XJs1 and XJs2 were most closely related to CS2-sat and CS1-sat, respectively, with 98.9% and 98.5% nucleotide sequence identity. Phylogenetic analysis of nucleotide sequence and deduced amino acid sequence of coat protein gene revealed that XJ1 and XJ2 belong to subgroup IB but there exist some variation between them. Parallel analyses of nucleotide sequence of XJsl and XJs2 suggested that these two satellite RNAs probably originated from China.

A Chlorophyll-Less Barley Mutant “NYB” Is Insensitive to Water Stress

“NYB” is a chlorophyll-less barley mutant, which grows relatively slow and unhealthily. The effects of water stress on photosystem II (PSII) of NYB and its wild type (WT) were investigated. Unexpected results indicated that the mutant was more resistant to water stress, because: PSII core proteins D1, D2 and LHCII declined more in WT than in NYB under water stress, and the corresponding psbA, psbD and cab mRNAs also decreased more dramatically in WT; CO2 assimilation, stomatal conductance, maximum efficiency of PSII photochemistry (Fv/Fm), efficiency of excitation energy capture by open PSII reaction centres (Fv’/Fm’), quantum yield of PSII electron transport (φPSII) and DCIP photoreduction in NYB were less sensitive to water stress than in WT, although the non-photochemical quenching coefficient (qN) and the photochemical quenching coefficient (qP) were almost the same in NYB and WT. Effective chlorophyll utilization and improved PSII protein formation in the mutant may be the reason for the enhanced stress resistance. Other possible mechanisms are also discussed.

Nuclear-Localized Plastid DNA Fragments in Protozoa, Metazoa and Fungi

We analyzed nuclear-localized plastid-like DNA (nupDNA) fragments in protozoa, metazoa and fungi. Most eukaryotes that do not have plastids contain 40-5000 bp nupDNAs in their nuclear genomes. These nupDNA fragments are mainly derived from repeated regions of plastids and distribute through the whole genomes. A majority of nupDNA fragments is located on coding regions with very important functions. Similar to plastids, these nupDNAs most possibly originate from cyanobacteria. Analysis of them suggests that through millions of years of universal endosymbiosis and gene transfer they may have occurred in ancient protists before divergence of plants and animals/fungi, and some transferred fragments have been reserved till now even in modern mammals.