Complete plastome sequence of Dracaena cambodiana (Asparagaceae): a species considered "Vulnerable" in Southeast Asia.

Dracaena cambodiana (Asparagaceae) is a treelike plant ranging from 3 to 10 m tall. It grows in low-elevation forests (0-300 m) having dry and sandy soils. It is distributed in Southern Hainan Island in China and other Southeast Asian countries (e.g. Cambodia, Laos, Thailand and Vietnam). The dried resin can be used medicinally as a substitute for that of Dracaena cochinchinensis. It has been ranked as a Vulnerable (VU) species in China. Here we report and characterize the complete plastid genome sequence of D. cambodiana. The complete plastome is 156,697 bp in length. It contains the typical structure and gene content of angiosperm plastomes, including two Inverted Repeat (IR) regions of 26,526 bp, a Large Single-Copy (LSC) region of 84,988 bp and a Small Single-Copy (SSC) region of 18,657 bp. The plastome contains 113 genes, consisting of 76 unique protein-coding genes, 30 unique tRNA genes, four unique rRNA genes and three pseudogenes (i.e. matK, infA, ndhF). The overall A/T content in the plastome of D. cambodiana is 62.4%. We performed phylogenetic analyses using the entire plastome, including spacers, introns, etc., and we determined that D. cambodiana and Maianthemum bicolor were closely related. The complete plastome sequence of D. cambodiana will provide a useful resource for the conservation genetics of this species as well as for phylogenetic studies in Asparagales.

Variation of Soil Bacterial Communities in a Chronosequence of Rubber Tree (Hevea brasiliensis) Plantations.

Regarding rubber tree plantations, researchers lack a basic understanding of soil microbial communities; specifically, little is known about whether or not soil microbial variation is correlated with succession in these plantations. In this paper, we used high-throughput sequencing of the 16S rRNA gene to investigate the diversity and composition of the soil bacterial communities in a chronosequence of rubber tree plantations that were 5, 10, 13, 18, 25, and 30 years old. We determined that: (1) Soil bacterial diversity and composition show changes over the succession stages of rubber tree plantations. The diversity of soil bacteria were highest in 10, 13, and 18 year-old rubber tree plantations, followed by 30 year-old rubber tree plantations, whereas 5 and 25 year-old rubber tree plantations had the lowest values for diversity. A total of 438,870 16S rDNA sequences were detected in 18 soil samples from six rubber tree plantations, found in 28 phyla, 66 classes, 139 orders, 245 families, 355 genera, and 645 species, with 1.01% sequences from unclassified bacteria. The dominant phyla were Acidobacteria, Proteobacteria, Chloroflexi, Actinobacteria, and Verrucomicrobia (relative abundance large than 3%). There were differences in soil bacterial communities among different succession stages of rubber tree plantation. (2) Soil bacteria diversity and composition in the different stages was closely related to pH, vegetation, soil nutrient, and altitude, of which pH, and vegetation were the main drivers.

An antibody against a conserved C-terminal consensus motif from plant alternative oxidase (AOX) isoforms 1 and 2 label plastids in the explosive dwarf mistletoe (Arceuthobium americanum, Santalaceae) fruit exocarp.

Dwarf mistletoes, genus Arceuthobium (Santalaceae), are parasitic angiosperms that spread their seeds by an explosive process. As gentle heating triggers discharge in the lab, we wondered if thermogenesis (endogenous heat production) is associated with dispersal. Thermogenesis occurs in many plants and is enabled by mitochondrial alternative oxidase (AOX) activity. The purpose of this study was to probe Arceuthobium americanum fruit (including seed tissues) collected over a 10-week period with an anti-AOX antibody/gold-labeled secondary antibody to determine if AOX could be localized in situ, and if so, quantitatively assess whether label distribution changed during development; immunochemical results were evaluated with Western blotting. No label could be detected in the mitochondria of any fruit or seed tissue, but was observed in fruit exocarp plastids of samples collected in the last 2 weeks of study; plastids collected in week 10 had significantly more label than week 9 (p = 0.002). Western blotting of whole fruit and mitochondrial proteins revealed a signal at 30-36 kD, suggestive of AOX, while blots of whole fruit (but not mitochondrial fraction) proteins showed a second band at 40-45 kD, in agreement with plastid terminal oxidases (PTOXs). AOX enzymes are likely present in the A. americanum fruit, even though they were not labeled in mitochondria. The results strongly indicate that the anti-AOX antibody was labeling PTOX in plastids, probably at a C-terminal region conserved in both enzymes. PTOX in plastids may be involved in fruit ripening, although a role for PTOX in thermogenesis cannot be eliminated.

Fungal endophytes of the obligate parasitic dwarf mistletoe Arceuthobium americanum (Santalaceae) act antagonistically in vitro against the native fungal pathogen Cladosporium (Davidiellaceae) of their host.

PREMISE OF THE STUDY: Endophytic fungi likely occur in all plants, yet little is known about those of parasitic plants, despite their potential to influence parasite success. Arceuthobium americanum is a parasitic angiosperm that greatly compromises the North American timber industry. We hypothesized that (1) A. americanum hosts fungal endophytes, and (2) these endophytes help A. americanum resist infection by fungal pathogens. • METHODS: Healthy A. americanum stem and fruit tissues were differentially stained for cellulose and chitin and visualized using fluorescence microscopy. Stem sections (sterilized vs. unsterilized) and seeds were incubated on agar plates to cultivate fungi, both to extract DNA for ITS rDNA sequencing and to observe interactions with native fungi from unsterilized specimens. • KEY RESULTS: Aside from xylem vessel elements, fungal structures were observed in all tissues, including those of the embryo. The ITS sequences of fungi cultured from internal tissues closely matched those of the known endophytes Phoma, Sydowia, and Phacidiopycnis, while those of surface organisms closely matched Cladosporium spp. Cultured fungi from internal tissues (putative endophytes) inhibited the growth of the surface organisms without affecting the other endophytes. • CONCLUSION: Fungal communities are established in A. americanum stems as well as in fruits and seeds, suggesting vertical transmission. These internally derived fungi act antagonistically toward fungi with pathogenic tendencies. As such, native mistletoe endophytes might protect A. americanum against fungal pathogens in nature. In the future, manipulation of endophytes might be a component of mistletoe control programs.

Bioactivity and the first transmission electron microscopy immunogold studies of short de novo-designed antimicrobial peptides.

In light of the era of microbial drug resistance, the current study aimed to better understand the relationships between sequence, higher-order structure, and mechanism of action for five designed peptides against multidrug-resistant (MDR) pathogens. All peptides studied were 15 residues long, were polycationic, adopted alpha-helical structures within hydrophobic environments (excluding the d-amino acid-substituted peptide MA-d), and contained N-terminal glycine residues, a novel antimicrobial peptide (AMP) design principle. Increasing hydrophobicity enhanced MICs (≤500 µg/ml to ≤7.4 µg/ml) without significantly increasing hemolytic activity (18% maximum hemolysis at 3,400 µg/ml). To the best of our knowledge, this is the first study to have successfully adapted and used a transmission electron microscopy (TEM) immunogold method to investigate the mechanism of action of short (∼15 residues long) AMPs within bacteria. We propose a "floodgate" mechanism to possibly explain membrane deformation and the relative absence of membrane-associated peptides 10 h into incubation.

Anatomy of leaf abscission in the Amur honeysuckle (Lonicera maackii, Caprifoliaceae): a scanning electron microscopy study.

Lonicera maackii (Rupr.) Maxim. (Amur honeysuckle) is native to Asia and an important ornamental in China. However, the anatomy of leaf abscission (shedding) in L. maackii had not been studied previously. Such work is needed not only because knowledge of the leaf abscission process is important for a horticultural species like L. maackii but also because leaf abscission is probably the least understood abscission process, as it occurs so rapidly. Therefore, our objective was to use scanning electron microscopy (SEM) to examine the progression of leaf abscission in L. maackii at the cellular level. L. maackii branches with leaves were regularly collected in Beijing, China over the 2-month period in which leaves abscise, and examined with SEM. We found that, unlike in model species, the cortex is involved in abscission, forming an "abaxial gap." We discovered that there is no discrete abscission zone prior to the onset of abscission and that no cell divisions precede abscission. An abscission zone did become evident well after the abscission process had begun, but its cells were enlarged, not constricted as in typical abscission zones. In the abaxial gap, intact cells separated at their middle lamella, but in the abscission zone, cell separation involved the entire wall, which is not typical. We did observe expected mechanical fission of vascular tissues. While the leaf abscission process we observed in L. maackii has similarities with model systems, aspects deviate from the expected.