Disposable stainless steel working electrodes for sensitive and simultaneous detection of indole-3-acetic acid and salicylic acid in Arabidopsis thaliana leaves under biotic stresses.

The detection of phytohormones in real time has attracted increasing attention because of their critical roles in regulating the development and signaling of plants, especially in defense against biotic stresses. Herein, stainless steel sheet electrodes modified with carbon cement were coupled with paper-based analysis devices for direct and simultaneous detection of salicylic acid (SA) and indole-3-acetic acid (IAA) in plants. We demonstrated that the excellent conductivity of stainless steel sheet electrodes enabled us to simultaneously differentiate IAA and SA at a level of 10 nM. With our approach, the content of IAA and SA in Arabidopsis thaliana leaves infected or not infected with Pst DC3000 could be rapidly quantified at the same time. Our experimental results on differentiation of IAA and SA at different time points showed that there were antagonistic interactions between the IAA and SA after infection of Arabidopsis leaves with Pst DC3000. By offering a cost-effective approach for rapid and sensitive detection of IAA and SA, this study suggests that electrochemical detection can be used in the study and development of precision agriculture technology.

Genome-Wide Characterization and Expression Analysis of Fatty acid Desaturase Gene Family in Poplar.

Fatty acid desaturases (FADs) modulate carbon-carbon single bonds to form carbon-carbon double bonds in acyl chains, leading to unsaturated fatty acids (UFAs) that have vital roles in plant growth and development and their response to environmental stresses. In this study, a total of 23 Populus trichocarpaFAD (PtFAD) candidates were identified from the poplar genome and clustered into seven clades, including FAB2, FAD2, FAD3/7/8, FAD5, FAD6, DSD, and SLD. The exon-intron compositions and conserved motifs of the PtFADs, clustered into the same clade, were considerably conserved. It was found that segmental duplication events are predominantly attributable to the PtFAD gene family expansion. Several hormone- and stress-responsive elements in the PtFAD promoters implied that the expression of the PtFAD members was complicatedly regulated. A gene expression pattern analysis revealed that some PtFAD mRNA levels were significantly induced by abiotic stress. An interaction proteins and gene ontology (GO) analysis indicated that the PtFADs are closely associated with the UFAs biosynthesis. In addition, the UFA contents in poplars were significantly changed under drought and salt stresses, especially the ratio of linoleic and linolenic acids. The integration of the PtFAD expression patterns and UFA contents showed that the abiotic stress-induced PtFAD3/7/8 members mediating the conversion of linoleic and linolenic acids play vital roles in response to osmotic stress. This study highlights the profiles and functions of the PtFADs and identifies some valuable genes for forest improvements.

Diversification in Qinghai-Tibet Plateau: Orchidinae (Orchidaceae) clades exhibiting pre-adaptations play critical role.

We explore the origins of the extraordinary plant diversity in the Qinghai-Tibetan Plateau (QTP) using Orchidinae (Orchidaceae) as a model. Our results indicate that six major clades in Orchidinae exhibited substantial variation in the temporal and spatial sequence of diversification. Our time-calibrated phylogenetic model suggests that the species-richness of Orchidinae arose through a combination of in situ diversification, colonisation, and local recruitment. There are multiple origins of species-richness of Orchidinae in the QTP, and pre-adaptations in clades from North Temperate and alpine regions were crucial for in situ diversification. The geographic analysis identified 29 dispersals from Asia, Africa and Europe into the QTP and 15 dispersals out. Most endemic species of Orchidinae evolved within the past six million years.

The evolution of floral deception in Epipactis veratrifolia (Orchidaceae): from indirect defense to pollination.

BACKGROUND: It is estimated that floral deception has evolved in at least 7500 species of angiosperms, of which two thirds are orchids. Epipactis veratrifolia (Orchidaceae) is a model system of aphid mimicry as aphidophagous hoverflies lay eggs on false brood sites on their flowers. To understand the evolutionary ecology of floral deception, we investigated the pollination biology of E. veratrifolia across 10 populations in the Eastern Himalayas. We reconstructed the phylogeny of Epipactis and mapped the known pollination systems of previously studied species onto the tree. RESULTS: Some inflorescences of E. veratrifolia were so infested with aphids while they were still in bud that the some larvae of hoverflies developed to the third instar while flower buds opened. This indicated that adult female hoverflies were partly rewarded for oviposition. Although flowers failed to secrete nectar, they mimicked both alarm pheromones and aphid coloring of to attract female hoverflies as their exclusive pollinators. Phylogenetic mapping indicate that pollination by aphidophagous hoverflies is likely an ancestral condition in the genus Epipactis. We suggest that the biological interaction of aphid (prey), orchid (primary producer) and hoverfly (predator) may represent an intermediate stage between mutualism and deception in the evolution of pollination-by-deceit in E. veratrifolia. CONCLUSIONS: Our analyses indicate that this intermediate stage may be used as a model system to interpret the origin of oviposition (brood site) mimicry in Epipactis. We propose the hypothesis that some deceptive pollination systems evolved directly from earlier (partly) mutualistic systems that maintained the fidelity of the original pollinator(s) even though rewards (nectar/ brood site) were lost.

Electrochemical sensors for plant signaling molecules.

Plant signaling molecules can be divided into plant messenger signaling molecules (such as calcium ions, hydrogen peroxide, Nitric oxide) and plant hormone signaling molecules (such as auxin (mainly indole-3-acetic acid or IAA), salicylic acid, abscisic acid, cytokinin, jasmonic acid or methyl jasmonate, gibberellins, brassinosteroids, strigolactone, and ethylene), which play crucial roles in regulating plant growth and development, and response to the environment. Due to the important roles of the plant signaling molecules in the plants, many methods were developed to detect them. The development of in-situ and real-time detection of plant signaling molecules and field-deployable sensors will be a key breakthrough for botanical research and agricultural technology. Electrochemical methods provide convenient methods for in-situ and real-time detection of plant signaling molecules in plants because of their easy operation, high sensitivity, and high selectivity. This article comprehensively reviews the research on electrochemical detection of plant signaling molecules reported in the past decade, which summarizes the various types electrodes of electrochemical sensors and the applications of multiple nanomaterials to enhance electrode detection selectivity and sensitivity. This review also provides examples to introduce the current research trends in electrochemical detection, and highlights the applicability and innovation of electrochemical sensors such as miniaturization, non-invasive, long-term stability, integration, automation, and intelligence in the future. In all, the electrochemical sensors can realize in-situ, real-time and intelligent acquisition of dynamic changes in plant signaling molecules in plants, which is of great significance for promoting basic research in botany and the development of intelligent agriculture.

Taxonomic notes on the genus Deutzia (Hydrangeaceae) from Central China.

Based on examination of syntype specimens deposited at P, the lectotype for the name Deutziasetchuenensis Franch. is designated here. By consulting literature and specimen records, the type locality of D.setchuenensisvar.longidentata Rehder, 'Chin-Ting shan' in the protologue is likely a misspelling of 'Chiuting shan' which is now called Jiuding shan located in southern Mao county, Sichuan province. In addition, a new variety, Deutziasetchuenensisvar.macrocarpa Q.L.Gan, Z.Y.Li & S.Z.Xu from western Hubei, Central China, is described and illustrated. It differs from other varieties of D.setchuenensis Franch. by the orange anthers, broader outer filaments, obtuse inner filaments, and larger fruits.

Astragalusbashanensis (Leguminosae), a new species from Central China.

A new species Astragalusbashanensis, from western Hubei Province, Central China is described and illustrated. The new species is morphologically similar to Astragalussinicus and A.wulingensis, but differs from both by its spreading pubescent indumentum on stems and petioles, longer petioles, white bracts, whitish or yellow corolla, longer claw of the keel-petal, hairy pods and smaller seeds.

Veronicahongii (Plantaginaceae), a new species from Central China.

A new species Veronicahongii, from western Hubei Province, Central China is described and illustrated. The species is morphologically similar to V.henryi Yamazaki, but mainly differs in the glabrous plant, except pedicels, broadly ovate leaf blades, glandular-pubescent pedicels, obovate calyx lobes, smaller corolla, broadly ovate capsule and much smaller seeds.

A new species of Prunus subgen. Cerasus from Central China.

A new species, Prunuswangii Q.L.Gan, Z.Y.Li & S.Z.Xu from western Hubei, Central China is described and illustrated. It is morphologically similar to P.clarofolia Schneid. and P.pseudocerasus Lindl., but differs in larger height, nearly erect branches, densely and horizontally arranged lenticels, straight lateral veins of leaves, persistent brownish bracts, reflexed and entire calyx lobes, 2-lobed petals with narrowly triangular sinus, earlier flowering and broadly ellipsoid fruits. Furthermore, P.wangii blooms in late February and the colour of flower changes with time, which makes it possible to be a new breeding material for ornamental cherry with early spring blooms.

Comprehensive Analysis of NAC Genes Reveals Differential Expression Patterns in Response to Pst DC3000 and Their Overlapping Expression Pattern during PTI and ETI in Tomato.

NAC (NAM/ATAF/CUC) transcription factors belong to a unique gene family in plants, which play vital roles in regulating diverse biological processes, including growth, development, senescence, and in response to biotic and abiotic stresses. Tomato (Solanum lycopersicum), as the most highly valued vegetable and fruit crop worldwide, is constantly attacked by Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), causing huge losses in production. Thus, it is essential to conduct a comprehensive identification of the SlNAC genes involved in response to Pst DC3000 in tomato. In this study, a complete overview of this gene family in tomato is presented, including genome localization, protein domain architectures, physical and chemical features, and nuclear location score. Phylogenetic analysis identified 20 SlNAC genes as putative stress-responsive genes, named SSlNAC 1-20. Expression profiles analysis revealed that 18 of these 20 SSlNAC genes were significantly induced in defense response to Pst DC3000 stress. Furthermore, the RNA-seq data were mined and analyzed, and the results revealed the expression pattern of the 20 SSlNAC genes in response to Pst DC3000 during the PTI and ETI. Among them, SSlNAC3, SSlNAC4, SSlNAC7, SSlNAC8, SSlNAC12, SSlNAC17, and SSlNAC19 were up-regulated against Pst DC3000 during PTI and ETI, which suggested that these genes may participate in both the PTI and ETI pathway during the interaction between tomato and Pst DC3000. In addition, SSlNAC genes induced by exogenous hormones, including indole-3-acetic acid (IAA), abscisic acid (ABA), salicylic acid (SA), and methyl jasmonic acid (MeJA), were also recovered. These results implied that SSlNAC genes may participate in the Pst DC3000 stress response by multiple regulatory pathways of the phytohormones. In all, this study provides important clues for further functional analysis and of the regulatory mechanism of SSlNAC genes under Pst DC3000 stress.

Disposable Stainless-Steel Wire-Based Electrochemical Microsensor for In Vivo Continuous Monitoring of Hydrogen Peroxide in Vein of Tomato Leaf.

As one of the pivotal signal molecules, hydrogen peroxide (H2O2) has been demonstrated to play important roles in many physiological processes of plants. Continuous monitoring of H2O2 in vivo could help understand its regulation mechanism more clearly. In this study, a disposable electrochemical microsensor for H2O2 was developed. This microsensor consists of three parts: low-cost stainless-steel wire with a diameter of 0.1 mm modified by gold nanoparticles (disposable working electrode), an untreated platinum wire with a diameter of 0.1 mm (counter electrode), and an Ag/AgCl wire with a diameter of 0.1 mm (reference electrode), respectively. The microsensor could detect H2O2 in levels from 10 to 1000 µM and exhibited excellent selectivity. On this basis, the dynamic change in H2O2 in the vein of tomato leaf under high salinity was continuously monitored in vivo. The results showed that the production of H2O2 could be induced by high salinity within two hours. This study suggests that the disposable electrochemical microsensor not only suits continuously detecting H2O2 in microscopic plant tissue in vivo but also reduces the damage to plants. Overall, our strategy will help to pave the foundation for further investigation of the generation, transportation, and elimination mechanism of H2O2 in plants.

The complete chloroplast genome sequence of Catalpa fargesii (Bignoniaceae), a species endemic to China.

Catalpa fargesii Bur. is endemic to China. Its complete chloroplast genome sequence was firstly reported in this study. The whole chloroplast genome of this species was 157765 bp in length including a pair of inverted repeat (IR, 30252 bp) regions separated by a small single copy (SSC, 12662 bp) and a large single copy (LSC, 84599 bp). The genome consisted of 134 genes, including 89 protein-coding genes, 8 rRNA and 37 tRNA genes. The phylogenetic analysis strongly supported that C. fargesii was closely related to C. fargesii f. duclouxii and C. bungei.

Flueggea acicularis (Phyllanthaceae), a narrow endemic species rediscovered in central China.

Flueggea acicularis (Phyllanthaceae) is endemic to the karst region of central China. Male specimens of this species were first collected in 1908. In 1989, female plants of F. acicularis were found for the first time, but misidentified as a new species. Throughout this period the male plants of F. acicularis were mismatched with female plants of other species, and male plants had not been collected since 1908. Then, in March, 2009, the authors rediscovered a wild population of F. acicularis consisting of both male and female plants in Wuxi county, Chongqing municipality, China. Based on field investigation and examination of specimens, we matched the correct female and male plants of this species for the first time since its initial publication a century ago. A complete and accurate morphological description, distribution, habitat and phenology of this species are also provided. Furthermore, the conservation status of F. acicularis is assessed as "Near Threatened" (NT) according to the IUCN Red List criteria.

The complete chloroplast genome sequence of Catalpa fargesii f. duclouxii (Bignoniaceae).

The complete chloroplast genome sequence of Catalpa fargesii f. duclouxii C.A. May was firstly assembled and analyzed in this study. The whole genome of this species was 158164 bp in length, with a typical quadripartite structure. The large single copy (LSC) was 83986 bp, the small single copy (SSC) was 12660 bp, and both of the two inverted repeats (IRs) were 30259 bp, respectively. A total of 134 genes in the chloroplast genome were annotated, including 87 protein-coding genes, 8 ribosomal RNA (rRNA) genes, and 39 transfer RNA (tRNA) genes. The phylogenetic analysis showed that C. fargesii f. duclouxii was highly clustered with C. bungei.

DNA barcoding of invasive plants in China: A resource for identifying invasive plants.

Invasive plants have aroused attention globally for causing ecological damage and having a negative impact on the economy and human health. However, it can be extremely challenging to rapidly and accurately identify invasive plants based on morphology because they are an assemblage of many different families and many plant materials lack sufficient diagnostic characteristics during border inspections. It is therefore urgent to evaluate candidate loci and build a reliable genetic library to prevent invasive plants from entering China. In this study, five common single markers (ITS, ITS2, matK, rbcL and trnH-psbA) were evaluated using 634 species (including 469 invasive plant species in China, 10 new records to China, 16 potentially invasive plant species around the world but not introduced into China yet and 139 plant species native to China) based on three different methods. Our results indicated that ITS2 displayed largest intra- and interspecific divergence (1.72% and 91.46%). Based on NJ tree method, ITS2, ITS, matK, rbcL and trnH-psbA provided 76.84%, 76.5%, 63.21%, 52.86% and 50.68% discrimination rates, respectively. The combination of ITS + matK performed best and provided 91.03% discriminatory power, followed by ITS2 + matK (85.78%). For identifying unknown individuals, ITS + matK had 100% correct identification rate based on our database, followed by ITS/ITS2 (both 93.33%) and ITS2 + matK (91.67%). Thus, we propose ITS/ITS2 + matK as the most suitable barcode for invasive plants in China. This study also demonstrated that DNA barcoding is an efficient tool for identifying invasive species.

Evaluation of the DNA barcodes in Dendrobium (Orchidaceae) from mainland Asia.

DNA barcoding has been proposed to be one of the most promising tools for accurate and rapid identification of taxa. However, few publications have evaluated the efficiency of DNA barcoding for the large genera of flowering plants. Dendrobium, one of the largest genera of flowering plants, contains many species that are important in horticulture, medicine and biodiversity conservation. Besides, Dendrobium is a notoriously difficult group to identify. DNA barcoding was expected to be a supplementary means for species identification, conservation and future studies in Dendrobium. We assessed the power of 11 candidate barcodes on the basis of 1,698 accessions of 184 Dendrobium species obtained primarily from mainland Asia. Our results indicated that five single barcodes, i.e., ITS, ITS2, matK, rbcL and trnH-psbA, can be easily amplified and sequenced with the currently established primers. Four barcodes, ITS, ITS2, ITS+matK, and ITS2+matK, have distinct barcoding gaps. ITS+matK was the optimal barcode based on all evaluation methods. Furthermore, the efficiency of ITS+matK was verified in four other large genera including Ficus, Lysimachia, Paphiopedilum, and Pedicularis in this study. Therefore, we tentatively recommend the combination of ITS+matK as a core DNA barcode for large flowering plant genera.