Nitric oxide is involved in the regulation of guard mother cell division by inhibiting the synthesis of ACC.

A stoma forms by a series of asymmetric divisions of stomatal lineage precursor cell and the terminal division of a guard mother cell (GMC). GMC division is restricted to once through genetic regulation mechanisms. Here, we show that nitric oxide (NO) is involved in the regulation of the GMC division. NO donor treatment results in the formation of single guard cells (SGCs). SGCs are also produced in plants that accumulate high NO, whereas clustered guard cells (GCs) appear in plants with low NO accumulation. NO treatment promotes the formation of SGCs in the stomatal signalling mutants sdd1, epf1 epf2, tmm1, erl1 erl2 and er erl1 erl2, reduces the cell number per stomatal cluster in the fama-1 and flp1 myb88, but has no effect on stomatal of cdkb1;1 cyca2;234. Aminocyclopropane-1-carboxylic acid (ACC), a positive regulator of GMC division, reduces the NO-induced SGC formation. Further investigation found NO inhibits ACC synthesis by repressing the expression of several ACC SYNTHASE (ACS) genes, and in turn ACC represses NO accumulation by promoting the expression of HEMOGLOBIN 1 (HB1) encoding a NO scavenger. This work shows NO plays a role in the regulation of GMC division by modulating ACC accumulation in the Arabidopsis cotyledon.

Plastome structure, evolution, and phylogeny of Selaginella.

As one of the earliest land plant lineages, Selaginella is important for studying land plant evolution. It is the largest genus of lycophytes containing 700-800 species. Some unique characters of Selaginella plastomes have been reported, but based only on 20 species. There have been no plastome phylogenies of Selaginella based on a relatively large sampling, and no efforts have been made to resolve the phylogeny of the enigmatic Sinensis group whose relationships have been unclear based on small datasets. Here we investigated the structures of 59 plastomes representing 51 species covering all six subgenera and 18 sections of Selaginella except two sections and including the intriguing Sinensis group for the first time. Our major results include: (1) the plastome size of Selaginella ranges tremendously from 78,492 bp to 187,632 bp; (2) there are numerous gene losses in Selaginella comparing with other lycophytes, Isoëtaceae and Lycopodiaceae; (3) the gene contents and plastome structures in Selaginella vary lineage-specifically and all infrageneric taxa are well supported in the plastome phylogeny; (4) the ndh gene family tends to lose or pseudogenize in those species with DR structure and without other short or medium repeats; (5) the short and medium repeat regions in SC mediate many conformations causing diverse and complex plastome structures, and six new conformations are discovered; (6) forty-eight species sampled have high GC content (>50%) but three species in the Sinensis group have âˆ¼ 30% GC content in plastomes, similar to most vascular plants; (7) the Sinensis group is monophyletic, includes at least two subgroups, and has the smallest plastomes in land plants except some parasitic plants, and their plastomes do not contain any tRNAs; (8) the younger lineages in Selaginella tend to have higher GC content, whereas the older lineages tend to have lower GC content; and (9) because of incomplete genomic data and abnormal structures or some unknown reasons, even the concatenated plastomes could not well resolve the phylogenetic relationships in Selaginella with confidence, highlighting the difficulty in resolving the phylogeny and evolution of this particularly important land plant lineage.

Molecular cloning and characterization of the glutathione reductase gene from Stipa purpurea.

Reactive oxygen species (ROS) are a key factor in abiotic stresses; excess ROS is harmful to plants. Glutathione reductase (GR) plays an important role in scavenging ROS in plants. Here, a GR gene, named SpGR, was cloned from Stipa purpurea and characterized. The full-length open reading frame was 1497 bp, encoding 498 amino acids. Subcellular localization analysis indicated that SpGR was localized to both the plasma membrane and nucleus. The expression of SpGR was induced by cold, salt, and drought stresses. Functional analysis indicated that ectopic expression of SpGR in Arabidopsis thaliana resulted in greater tolerance to salt stress than that of wild-type plants, but no difference under cold or drought treatments. The results of GR activity and GSSG and GSH content analyses suggested that, under salt stress, transgenic plants produced more GR to reduce GSSG to GSH for scavenging ROS than wild-type plants. Therefore, SpGR may be a candidate gene for plants to resist abiotic stress.

A large-scale phylogeny of the lycophyte genus Selaginella (Selaginellaceae: Lycopodiopsida) based on plastid and nuclear loci.

The lycophyte genus Selaginella alone constitutes the family Selaginellaceae, the largest of the lycophyte families. The genus is estimated to contain 700-800 species distributed on all continents except Antarctica, with highest species diversity in tropical and subtropical regions. The monophyly of Selaginella in this broad sense has rarely been doubted, whereas its intrageneric classification has been notoriously contentious. Previous molecular studies were based on very sparse sampling of Selaginella (up to 62 species) and often used DNA sequence data from one genome. In the present study, DNA sequences of one plastid (rbcL) and one nuclear (ITS) locus from 394 accessions representing approximately 200 species of Selaginella worldwide were used to infer a phylogeny using maximum likelihood, Bayesian inference and maximum parsimony methods. The study identifies strongly supported major clades and well resolves relationships among them. Major results include: (i) six deep-level clades are discovered representing the deep splits of Selaginella; and (ii) 20 major clades representing 20 major evolutionary lineages are identified, which differ from one another in molecular, macro-morphological, ecological and spore features, and/or geographical distribution.

Selaginelladensiciliata (subg. Heterostachys, Selaginellaceae), a new spikemoss species from China based on morphological and molecular data.

A new species of spikemoss, Selaginelladensiciliata in S.subg.Heterostachyssect.Tetragonostachyae, China, is described from southeastern Xizang, based on morphological and molecular phylogenetic data. Morphologically, S.densiciliata is similar to S.repanda, S.subvaginata and S.vaginata, but the new species can be easily distinguished from them by having sterile leaves margins densely ciliate, symmetrical axillary leaves oblong ovate to ovate-triangular, and ovate dorsal leaves obviously carinate. Molecular phylogenetic analysis resolves S.densiciliata as sister to the clade comprised with S.vaginata and S.xipholepis, which confirms the recognition of the new species.

Transcriptome-Based Study on the Phylogeny and Hybridization of Marattialean Ferns (Marattiaceae).

Marattiaceae is a phylogenetically isolated family of tropical eusporangiate ferns including six genera with more than one-hundred species. In Marattiaceae, monophyly of genera has been well-supported phylogenetically. However, the phylogenetic relationships among them were elusive and controversial. Here, a dataset of 26 transcriptomes (including 11 newly generated) were used to assess single-copy nuclear genes and to obtain the organelle gene sequences. Through phylotranscriptomic analysis, the phylogeny and hybridization events of Marattiaceae were explored and a robust phylogenomic framework for the evolution of Marattiaceae was provided. Using both concatenation- and coalescent-based phylogenies, the gene-tree discordance, incomplete lineage sorting (ILS) simulations, and network inference were examined. Except the low support with mitochondrial genes of Marattiaceae, nuclear genes and chloroplast genes strongly supported a sister relationship between Marattiaceae and leptosporangiate ferns. At the genus level, all phylogenetic analysis based on nuclear genes datasets recovered five genera in Marattiaceae as monophyletic with strong support. Danaea and Ptisana were the first two diverged clades in turn. Christensenia was a sister clade to the clade Marattia + Angiopteris s.l. In Angiopteris s.l., three clades (Angiopteris s.s., the Archangiopteris group, and An. sparsisora) were well identified with maximum support. The Archangiopteris group was derived from Angiopteris s.s. at ca. 18 Ma. The putative hybrid species An. sparsisora between Angiopteris s.s. and the Archangiopteris group was verified by the species network analyses and the maternal plastid genes. This study will improve our understanding for using the phylotranscriptomic method to explore phylogeny and investigate hybridization events for difficult taxa in ferns.

Carbon monoxide promotes stomatal initiation by regulating the expression of two EPF genes in Arabidopsis cotyledons.

The gaseous molecule carbon monoxide (CO) can freely pass through the cell membrane and participate in signal transduction in the cell to regulate physiological activities in plants. Here, we report that CO has a positive regulatory role in stomatal development. Exogenous CO donor CORM-2 [Tricarbonyldichlororuthenium (II) dimer] treatment resulted in an increase of stomatal index (SI) on the abaxial epidermis of cotyledons in wild-type, which can be reversed by the addition of the CO biosynthesis inhibitor ZnPPIX [Protoporphyrin IX zinc (II)]. Consistent with this result, mutation of the CO biosynthesis gene HY1 resulted in a decrease of SI in hy1-100 plants, while overexpression of HY1 led to an increase of SI. Further investigation revealed that CO acts upstream of SPCH and YDA in the stomatal development pathway, since the loss of function mutants spch-1 and yda-2 were insensitive to CORM-2. The expression of EPF2 was inhibited by CORM-2 treatment in wild type and is lower in hy1 than in wild-type plants. In contrast, the expression of STOMAGEN was promoted by CORM-2 treatment and is higher in HY1-overexpression lines. Loss of function mutants of both epf2 and stomagen are insensitive to CORM-2 treatment. These results indicated that CO positively regulates stomatal initiation and distribution by modulating the expression of EPF2 and STOMAGEN.

The complete chloroplast genome of Endangered species Aristolochia delavayi Franch. (Aristolochiaceae) in Southwestern China.

The complete chloroplast genome of Aristolochia delavayi was determined in this study. The chloroplast genome consists of 160,344 bp, with a typical circular structure including a pair of inverted repeats of 25,454 bp separated by a large single-copy region and a small single-copy region of 89,502 and 19,795 bp, respectively. The plastome contains 130 genes, including 85 protein-coding genes, eight rRNA genes, and 37 tRNA genes. Further phylogenetic analyses were conducted using 12 complete plastomes of Aristolochia. These data support a close relationship between Aristolochia delavayi and Aristolochia tubiflora.

[Cadmium Accumulation Characteristics of Four Herbs].

A pot-based experiment was conducted to study the Cd tolerance and accumulation characteristics of four invasive herbs (Galinsoga quadriradiata, Panicum dichotomiflorum, Setaria geniculata, and Lolium persicum) under exposures of 0 (T0), 5 (T5), 25 (T25), and 50 mg·kg-1 (T50) soil Cd concentrations to screen for potential Cd accumulators for phytoremediation. The results showed that the biomasses of both shoots and roots of G. quadriradiata had no significant changes compared to the control (T0) samples under all Cd treatments, whereas the biomass of the other three Poaceae species significantly decreased under the T25 or T50 treatment. The results indicate that G. quadriradiata had stronger Cd tolerance than the other three species. The Cd concentrations in the shoots and roots of the four herbs significantly increased with an increase in soil Cd concentrations, but the shoot bioconcentration factors (SBCF) of the four plant species significantly decreased under T5, T25, and T50 treatment. The SBCF of G. quadriradiata and P. dichotomiflorum were greater than 1 whereas those of S. geniculata and L. persicum were lower than 1. The translocation factors (TF) of G. quadriradiata were 0.93, 0.73, and 1.04 under T5, T25, and T50 treatment, respectively, which were significantly higher than those of the other three plants under the same soil Cd concentration. In addition, both the total Cd and shoot Cd contents of G. quadriradiata were notably higher than in the other three species under the same Cd treatment. Moreover, 90% of the Cd in G. quadriradiata could be transferred aboveground, which was significantly higher than for the other three plants. Based on our comprehensive comparison of Cd tolerance and accumulation capacity, we suggest that G. quadriradiata is a high-Cd accumulator with considerable phytoremediation potential.