Symbiosis between Dendrobium catenatum protocorms and Serendipita indica involves the plant hypoxia response pathway.

Mycorrhizae are ubiquitous symbioses established between fungi and plant roots. Orchids, in particular, require compatible mycorrhizal fungi for seed germination and protocorm development. Unlike arbuscular mycorrhizal fungi, which have wide host ranges, orchid mycorrhizal fungi are often highly specific to their host orchids. However, the molecular mechanism of orchid mycorrhizal symbiosis is largely unknown compared to that of arbuscular mycorrhizal and rhizobial symbiosis. Here, we report that an endophytic Sebacinales fungus, Serendipita indica, promotes seed germination and the development of protocorms into plantlets in several epiphytic Epidendroideae orchid species (6 species in 2 genera), including Dendrobium catenatum, a critically endangered orchid with high medicinal value. Although plant-pathogen interaction and high meristematic activity can induce the hypoxic response in plants, it has been unclear whether interactions with beneficial fungi, especially mycorrhizal ones, also involve the hypoxic response. By studying the symbiotic relationship between D. catenatum and S. indica, we determined that hypoxia-responsive genes, such as those encoding alcohol dehydrogenase (ADH), are highly induced in symbiotic D. catenatum protocorms. In situ hybridization assay indicated that the ADH gene is predominantly expressed in the basal mycorrhizal region of symbiotic protocorms. Additionally, the ADH inhibitors puerarin and 4-methylpyrazole both decreased S. indica colonization in D. catenatum protocorms. Thus, our study reveals that S. indica is widely compatible with orchids and that ADH and its related hypoxia-responsive pathway are involved in establishing successful symbiotic relationships in germinating orchids.

MITOCHONDRIA: The dual function of the transient receptor potential melastatin 2 channels from cytomembrane to mitochondria.

Mitochondria are closely related to oxidative stress and play an important role in maintaining cell functional homeostasis and meeting cell energy demand. The transient receptor potential melastatin 2 (TRPM2) channel affects the occurrence and progression of diseases by regulating mitochondrial function. TRPM2 channel promotes Ca2+ influx to affect 18 kDa translocator protein (TSPO), mitochondrial membrane potential (MMP), reactive oxygen species (ROS), adenosine triphosphate (ATP) production, and mitochondrial autophagy. The mechanism of Ca2+ influx into the mitochondria by TRPM2 is abundant. Interestingly, the TRPM2 channel inhibits the production of mitochondrial ROS in cancer cells and promotes the production of mitochondrial ROS in normal cells, which induces cell death in normal cells but proliferation in cancer cells. TRPM2 can be a potential target for the treatment of various diseases due to its role as a molecular link between mitochondria and Ca2+ signals.

Regulatory Effects of Clock and Bmal1 on Circadian Rhythmic TLR Expression.

Circadian locomotor output cycles kaput (Clock) and brain and muscle ARNT-like 1 (Bmal1) are two core circadian clock genes. They form a heterodimer that can bind to the E-box element in the promoters of Period circadian protein (Per) and Cryptochrome (Cry) genes, thereby inducing the rhythmic expression of circadian clock control genes. Toll-like receptors (TLRs) are type I transmembrane proteins belonging to the pattern recognition receptor (PRR) family. They can recognize a variety of pathogens and play an important role in innate immunity and adaptive immune responses. Recent studies have found that the circadian clock is closely associated with the immune system. TLRs have a certain correlation with the circadian rhythms; Bmal1 seems to be the central mediator connecting the circadian clock and the immune system. Research on Bmal1 and TLRs has made some progress, but the specific relationship between TLRs and Bmal1 remains unclear. Understanding the relationship between TLRs and Clock/Bmal1 genes is increasingly important for basic research and clinical treatment.

Seed viability testing for research and conservation of epiphytic and terrestrial orchids.

BACKGROUND: Seed viability testing is essential in plant conservation and research. Seed viability testing determines the success of ex-situ conservation efforts, such as seed banking but commonly testing protocols of orchids lack consistency and accuracy, therefore, there is a need to select an appropriate and reliable viability test, especially when conducting comparative studies. Here, we evaluated the suitability of three seed viability tests, Evans blue test (EB), Fluorescein diacetate test (FDA) and Tetrazolium test (TTC), with and without sterilization, on seeds of 20 orchid species, which included five epiphytes and fifteen terrestrials, using both fresh seeds and seeds stored at - 18 ºC for 6 to 8 years. RESULTS: We found that sterilization and lifeform of seeds affected seed viability across all tests but the storage time was not an influential factor. Sterilization negatively affected seed viability under EB and FDA test conditions but increased the detection of viable seeds in the TTC test in both epiphytic and terrestrial species. The EB test, when administered without sterilization provided the highest viability results. Being non-enzymatic unlike TTC and FDA tests, as expected, the EB test was the most reliable with similar results between sterilized and not sterilized seeds for most epiphytic and terrestrial species as well as when compared between groups. CONCLUSIONS: The lifeform of the species and seed sterilization prior to testing are important influential factors in orchid seed viability testing. Since EB test was found to be reliable we recommend the EB test for seed viability assessment in orchids rather than the less reliable but commonly used TTC test, or the FDA test, which require more expensive and sophisticated instrumentation. Since storage time was not an influential factor in orchid seed viability testing, the recommendations of this study can be used for both fresh as well as long-term stored orchid seeds. This is helpful for research and especially for conservation measures such as seed banking. However, due to the species specificity of the bio-physiology of orchids, we call for comprehensive viability test assessment in the hyper diverse orchid family to be extended to a greater number of species to facilitate efficient conservation and research.

Transitions between the Terrestrial and Epiphytic Habit Drove the Evolution of Seed-Aerodynamic Traits in Orchids.

Orchids are globally distributed, a feature often attributed to their tiny dustlike seeds. They were ancestrally terrestrial but in the Eocene expanded into tree canopies, with some lineages later returning to the ground, providing an evolutionarily replicated system. Because seeds are released closer to the ground in terrestrial species than in epiphytic ones, seed traits in terrestrials may have been under selective pressure to increase seed dispersal efficiency. In this study, we test the expectations that seed airspace-a trait known to increase seed flotation time in the air-is (i) larger in terrestrial lineages and (ii) has increased following secondary returns to a terrestrial habit. We quantified and scored 20 seed traits in 121 species and carried out phylogenetically informed analyses. Results strongly support both expectations, suggesting that aerodynamic traits even in dust seeds are under selection to increase dispersal ability, following shifts in average release heights correlated with changes in habit.

Minireview: The Introduction of COVID-19 and Microvascular Disease-Introduction of COVID-19 and the Relationship between Its Receptor and Diabetic Vascular Complications.

In December of 2019, a novel coronavirus, which is SARS-CoV-2, broke out in the world and caused tremendous human and financial losses. According to a descriptive study by the relative hospital about the epidemiological and clinical features of 52 critically ill patients, the expert panel found that people with cardiovascular disease and diabetes comprise a large proportion of the patients with chronic disease. In this review, we discuss the structural biology of the SARS-CoV-2 in combination with the characteristics of its binding protein, ACE2, which is an important receptor in the cardiovascular system and may have potential relationships with various diabetic diseases. We hope we can provide useful recommendations for patients with diabetes after becoming infected by the virus or provide directions to doctors on treatment options.

Using In Situ Symbiotic Seed Germination to Restore Over-collected Medicinal Orchids in Southwest China.

Due to increasing demand for medicinal and horticultural uses, the Orchidaceae is in urgent need of innovative and novel propagation techniques that address both market demand and conservation. Traditionally, restoration techniques have been centered on ex situ asymbiotic or symbiotic seed germination techniques that are not cost-effective, have limited genetic potential and often result in low survival rates in the field. Here, we propose a novel in situ advanced restoration-friendly program for the endangered epiphytic orchid species Dendrobium devonianum, in which a series of in situ symbiotic seed germination trials base on conspecific fungal isolates were conducted at two sites in Yunnan Province, China. We found that percentage germination varied among treatments and locations; control treatments (no inoculum) did not germinate at both sites. We found that the optimal treatment, having the highest in situ seed germination rate (0.94-1.44%) with no significant variation among sites, supported a warm, moist and fixed site that allowed for light penetration. When accounting for seed density, percentage germination was highest (2.78-2.35%) at low densities and did not vary among locations for the treatment that supported optimal conditions. Similarly for the same treatment, seed germination ranged from 0.24 to 5.87% among seasons but also did vary among sites. This study reports on the cultivation and restoration of an endangered epiphytic orchid species by in situ symbiotic seed germination and is likely to have broad application to the horticulture and conservation of the Orchidaceae.

Orchid conservation in the biodiversity hotspot of southwestern China.

Xishuangbanna is on the northern margins of tropical Asia in southwestern China and has the largest area of tropical forest remaining in the country. It is in the Indo-Burma hotspot and contains 16% of China's vascular flora in <0.2% of the country's total area (19,690 km(2) ). Rapid expansion of monoculture crops in the last 20 years, particularly rubber, threatens this region's exceptional biodiversity. To understand the effects of land-use change and collection on orchid species diversity and determine protection priorities, we conducted systematic field surveys, observed markets, interviewed orchid collectors, and then determined the conservation status of all orchids. We identified 426 orchid species in 115 genera in Xishuangbanna: 31% of all orchid species that occur in China. Species richness was highest at 1000-1200 m elevation. Three orchid species were assessed as possibly extinct in the wild, 15 as critically endangered, 82 as endangered, 124 as vulnerable, 186 as least concern, and 16 as data deficient. Declines over 20 years in harvested species suggested over-collection was the major threat, and utility value (i.e., medicinal or ornamental value) was significantly related to endangerment. Expansion of rubber tree plantations was less of a threat to orchids than to other taxa because only 75 orchid species (17.6%) occurred below the 1000-m-elevation ceiling for rubber cultivation, and most of these (46) occurred in nature reserves. However, climate change is projected to lift this ceiling to around 1300 m by 2050, and the limited area at higher elevations reduces the potential for upslope range expansion. The Xishuangbanna Tropical Botanical Garden is committed to achieving zero plant extinctions in Xishuangbanna, and orchids are a high priority. Appropriate in and ex situ conservation strategies, including new protected areas and seed banking, have been developed for every threatened orchid species and are being implemented.

Investigation of the inclusion interaction of p-sulfonatocalix[6]arene with triamterene.

The inclusion complexation behavior of p-sulfonatocalix[6]arene (SCX6) with triamterene (TA) was investigated by fluorescence spectroscopy, UV-Vis spectroscopy, (1)H and 2D NMR, FT-IR, SEM and DSC. The results indicate that TA is able to form an inclusion complex with SCX6. The inclusion complex has a stoichiometry of 1:1 at pH 6.50. In addition, the water solubility of TA was obviously increased in the inclusion complex with SCX6.

Rain pollination provides reproductive assurance in a deceptive orchid.

BACKGROUND AND AIMS: Abiotic pollination by wind or water is well established in flowering plants. In some species pollination by rain splashes, a condition known as ombrophily, has been proposed as a floral strategy. However, evidence for this type of abiotic pollination has remained controversial and many reported cases have subsequently been shown to be false. This study investigates ombrophily in the deceptive orchid Acampe rigida to determine the mechanism by which this species is able to maintain high fecundity, despite flowering during the rainy season in south-west China when pollinators are scarce. METHODS: The floral mechanisms promoting rain pollination in A. rigida were observed and described in detail. Controlled pollination experiments and observations of floral visitors were conducted. A field experiment using rain shelters at 14 sites in Guangxi, south-west China, evaluated the contribution of rain pollination to fruit-set. KEY RESULTS: During rainfall, raindrops physically flicked away the anther cap exposing the pollinarium. Raindrops then caused pollinia to be ejected upwards with the strap-like stipe pulling them back and causing them to fall into the stigmatic cavity, resulting in self-pollination. Neither flower nor pollen function were damaged by water. Although A. rigida is self-compatible, it is incapable of autonomous self-pollination without the assistance of rain splashes. The results of the rain-sheltering experiment indicated that rain pollination contributed substantially to increasing fruit-set, although there was variation among sites in the intensity of this effect. CONCLUSIONS: A. rigida flowers during the rainy season, when pollinators are scarce, and ombrophily functions to provide reproductive assurance without compromising opportunities for outcrossing.

Microsatellite markers for Duperrea pavettifolia (Rubiaceae).

PREMISE OF THE STUDY: The development of microsatellite primers for Duperrea pavettifolia will be the foundation for mating system analysis and conservation research. METHODS AND RESULTS: Nineteen microsatellite markers were developed and characterized in two wild populations by using the Fast Isolation by AFLP of Sequences COntaining repeats (FIASCO) protocol. Polymorphisms were evaluated in 24 individuals from two natural populations. Eleven of these primers generated polymorphic loci. CONCLUSIONS: These microsatellite markers will be useful in future investigations into the population genetics and mating system of D. pavettifolia.