Pharbitis Nil Research Articles

Methane emission, nitrogen excretion, and energy partitioning in Hanwoo steers fed a typical TMR diet supplemented with Pharbitis nil seeds.

Two in vivo experiments were conducted to evaluate the potential of Pharbitis nil seeds (PA) as an anti-methanogenic additive to ruminant feed. In experiment 1, six Hanwoo steers (459.0 ± 25.8 kg) were fed either a total mixed ration (TMR; 32-d period) or TMR supplemented with PA at 5% dry matter (DM) intake (TMR-PA; 45-d period) for two consecutive periods. Fecal and urine outputs were measured in an apparent digestibility trial in both periods. Methane (CH4) yield and heat energy (HE) were measured using respiratory chambers equipped with gas analyzers. In experiment 2, five rumen cannulated Holstein steers (744 ± 35 kg) were fed the same TMR or TMR-PA diets for 40 days; rumen samples were collected at 0, 1.5, and 3 h after feeding on the last day of the feeding period. In experiment 1, although there were no differences (p > 0.05) in nutrients or gross energy intake (GEI) between the groups, an increase (p < 0.05) in the apparent digestibility of DM (9.1%) and neutral detergent fiber (22.9%) was observed in the TMR-PA fed Hanwoo steers. Pronounced decreases (p < 0.05) in CH4 (g/Kg DM; 17.1%) and urinary N excretion (% N intake; 7.6%) were observed in the TMR-PA group, leading to a 14.7% increase in metabolizable energy intake (% GEI). However, only a numerical increase (p > 0.05) in retained energy was observed due to the increase in HE loss. In experiment 2, a drastic decrease (p < 0.05) in rumen ammonia concentration (56.3%) associated with an increased (p = 0.091) rumen short-chain fatty acid concentration 1.5 h after feeding were observed in TMR-PA fed Holstein steers. A 26.6% increase (p < 0.05) in the propionate proportion during the treatment period clearly reflected a shift in the ruminal H2 sink after 3 h of feeding. A 40% reduction (p = 0.067) in the relative abundance of rumen protozoa Entodinium caudatum was also observed. It was concluded that PA could be a natural feed additive for CH4 and N emission abatement.

New isolate of sweet potato virus 2 from Ipomoea nil: molecular characterization, codon usage bias, and phylogenetic analysis based on complete genome

BackgroundViral diseases of sweet potatoes are causing severe crop losses worldwide. More than 30 viruses have been identified to infect sweet potatoes among which the sweet potato latent virus (SPLV), sweet potato mild speckling virus (SPMSV), sweet potato virus G (SPVG) and sweet potato virus 2 (SPV2) have been recognized as distinct species of the genus Potyvirus in the family Potyviridae. The sweet potato virus 2 (SPV2) is a primary pathogen affecting sweet potato crops.MethodsIn this study, we detected an SPV2 isolate (named SPV2-LN) in Ipomoea nil in China. The complete genomic sequence of SPV2-LN was obtained using sequencing of small RNAs, RT-PCR, and RACE amplification. The codon usage, phylogeny, recombination analysis and selective pressure analysis were assessed on the SPV2-LN genome.ResultsThe complete genome of SPV2-LN consisted of 10,606 nt (GenBank No. OR842902), encoding 3425 amino acids. There were 28 codons in the SPV2-LN genome with a relative synonymous codon usage (RSCU) value greater than 1, of which 21 end in A/U. Among the 12 proteins of SPV2, P3 and P3N-PIPO exhibited the highest variability in their amino acid sequences, while P1 was the most conserved, with an amino acid sequence identity of 87-95.3%. The phylogenetic analysis showed that 21 SPV2 isolates were clustered into four groups, and SPV2-LN was clustered together with isolate yu-17-47 (MK778808) in group IV. Recombination analysis indicated no major recombination sites in SPV2-LN. Selective pressure analysis showed dN/dS of the 12 proteins of SPV2 were less than 1, indicating that all were undergoing negative selection, except for P1N-PISPO.ConclusionThis study identified a sweet potato virus, SPV2-LN, in Ipomoea nil. Sequence identities and genome analysis showed high similarity between our isolate and a Chinese isolate, yu-17-47, isolated from sweet potato. These results will provide a theoretical basis for understanding the genetic evolution and viral spread of SPV2.

Efficacy of rhizobacteria in suppressing weed growth

The production of bioherbicides from isolates of deleterious rhizobacteria capable of suppressing plant growth is a sustainable method for weed management. The objective of this research was to evaluate whether rhizobacteria that potentially inhibit plant growth interfere with the growth of weeds. The experiment was carried out in a greenhouse in a completely randomized design with four replications. The treatments consisted of inoculating seeds of the weeds Bidens pilosa L., Senna occidentalis (L.) Link, and Ipomoea nil (L.) Roth with two distinct isolates of rhizobacteria (171 and 122), as well as cultivating these weeds without inoculation. At 50 days after sowing of the weeds, the height, stem diameter, shoot dry matter, and root dry matter were measured. Inoculation with isolates 171 and 122 did not influence the growth in height, diameter, shoot dry matter, or root dry matter of the weeds. It is concluded that rhizobacteria 171 and 122 do not have the potential to inhibit the growth of the weeds B. pilosa, S. occidentalis, and I. nil.

A chemical approach to extend flower longevity of Japanese morning glory via inhibition of master senescence regulator EPHEMERAL1.

Petal senescence in flowering plants is a type of programmed cell death with highly regulated onset and progression. A NAM/ATAF1,2/CUC2 transcription factor, EPHEMERAL1 (EPH1), has been identified as a key regulator of petal senescence in Japanese morning glory (Ipomoea nil). Here we used a novel chemical approach to delay petal senescence in Japanese morning glory by inhibiting the DNA-binding activity of EPH1. A cell-free high-throughput screening system and subsequent bioassays found two tetrafluorophthalimide-based compounds, Everlastin1 and Everlastin2, that inhibited the EPH1-DNA interaction and delayed petal senescence. The inhibitory mechanism was due to the suppression of EPH1 dimerization. RNA-sequencing analysis revealed that the chemical treatment strongly suppressed the expression of programmed cell death- and autophagy-related genes. These results suggest that a chemical approach targeting a transcription factor can regulate petal senescence.

Mycena citricolor isolates from diverse hosts in Costa Rica: Colony aspect, growth dynamics, reproduction in vitro and aggressiveness on Coffea arabica

AbstractThe basidiomycete fungus Mycena citricolor hosts on about 80 plant families. The asexual morph causes the disease known as American leaf spot of coffee (Coffea arabica). We isolated and characterized 15 isolates of M. citricolor collected from coffee plants, shade trees and weeds. Cultures on potato‐dextrose‐agar + yeast extract (PDAY) medium were kept for a week in complete darkness. The cultures were then exposed to light for 7 weeks and co‐cultivated with coffee foliar discs, to stimulate the production of gemmifers. We evaluated the speed of growth, appearance, colour of the mycelium and gemmifers production capacity. All isolates conformed to the typical cultural characteristics of M. citricolor, with beige/white thin and slightly aerial mycelium. The whitest isolate was from Persea americana, which had the slowest growth rate in vitro (6.4 mm/day), while the isolate from weed Kalanchoe pinnata was the fastest, with a speed of 9 mm/day. The most gemmifer‐producing isolates were those isolated from the coffee cultivar Catimor‐Costa Rica 95, Citharexylum donnell‐smithii and Cissus verticillata, with over 200 gemmifers per Petri dish. Isolates from Erythrina and Ipomoea nil produced &lt;50 gemmifers on average. The Composite Aggressiveness Index (CAI) was determined on 2‐year‐old detached coffee branches of the Caturra variety foliage. The least aggressive isolate was from Erythrina poeppigiana (CAI = 0) and the most aggressive was from Kalanchoe pinnata (CAI = 18.87). The isolate from the coffee cultivar Catimor‐Costa Rica 95 was more aggressive than that from the cultivar Caturra (CAI = 9.94 vs. CAI = 3.66). Moreover, successful infection occurred only when the apex of gemmae made direct contact with the coffee leaf when using the inoculum derived from C. verticillata and Anredera cordifolia. These results show the variability in the population of the pathogen. This is the first study that characterizes the growth, morphology, reproduction and aggressiveness of M. citricolor obtained from hosts other than coffee. In addition, it is the first to report the use of coffee foliar discs to induce gemmifer production in vitro and to assess and quantify the effect of gemma position when inoculating coffee leaves.

Flexible Resource Allocation-Efficient Water Use Strategies Facilitate Invasion of Invasive Vine Sicyos angulatus L.

The invasive vine Sicyos angulatus L. destroys the natural ecosystem of invaded areas. Understanding the differences in growth and development between S. angulatus and other plants is necessary to explore the invasion mechanisms of S. angulatus and implement appropriate prevention and control measures. Thus, this study compared the growth, photosynthesis, and root characteristics of invasive liana S. angulatus and other three vine plants, Ipomoea nil (L.) Roth, Ipomoea purpurea (L.), and Thladiantha dubia Bunge, at different growth stages: seedling, flowering, and fruiting. The results showed that the total biomass of S. angulatus in the fruiting stage was 3-6 times that of the other three plants, and the root biomass ratio and root-shoot ratio decreased throughout the growth stage. Throughout the growth stage, the total leaf area of S. angulatus was significantly higher than that of the other three plant types, and the specific leaf area of S. angulatus at the seedling and flowering stages was 2.5-3 and 1.4-3 times that of the other three plants, respectively. The photosynthetic rate, stomatal conductance, and transpiration rate of S. angulatus at the fruiting stage were significantly higher than those of the other three plants, and its water use efficiency was higher than that of the other three plants at the three growth stages, indicating its strong photosynthetic capacity. The root activity and root pressure of S. angulatus were also significantly higher than those of the other three plants at the seedling and flowering stages. These results show that S. angulatus flexibly allocates resources to its aboveground parts during the growth stage to ensure that the plant obtains the space necessary for its growth and development and that with the help of higher root pressure and root activity, S. angulatus can maintain higher photosynthesis and water use efficiency with fewer resources. Therefore, the prevention and control of S. angulatus requires a combination of aboveground and underground measures. Spraying conventional weedicide/herbicide and manually removing aboveground plants may lead to its resurgence.

ACRE, a class of AP2/ERF transcription factors, activates the expression of sweet potato ß-amylase and sporamin genes through the sugar-responsible element CMSRE-1

Sugars, synthesized by photosynthesis in source organs, are loaded and utilized as an energy source and carbon skeleton in sink organs, and also known to be important signal molecules regulating gene expression in higher plants. The expression of genes coding for sporamin and β-amylase, the two most abundant proteins in storage roots of sweet potato, is coordinately induced by sugars. We previously reported on the identification of the carbohydrate metabolic signal-responsible element-1 (CMSRE-1) essential for the sugar-responsible expression of two genes. However, transcription factors that bind to this sequence have not been identified. In this study, we performed yeast one-hybrid screening using the sugar-responsible minimal promoter region of the ß-amylase gene as bait and a library composed only transcription factor cDNAs of Arabidopsis. Two clones, named Activator protein binding to CMSRE-1 (ACRE), encoding AP2/ERF transcription factors were isolated. ACRE showed transactivation activity of the sugar-responsible minimal promoter in a CMSRE-1-dependent manner in Arabidopsis protoplasts. Electric mobility shift assay (EMSA) using recombinant proteins and transient co-expression assay in Arabidopsis protoplasts revealed that ACRE could actually act to the CMSRE-1. Among the DEHYDRATION -RESPONSIVE ELEMENT BINDING FACTOR (DREB) subfamily, almost all homologs including ACRE, could act on the DRE, while only three ACREs could act to the CMSRE-1. Moreover, ACRE-homologs of Japanese morning glory also have the same property of DNA-binding preference and transactivation activity through the CMSRE-1. These findings suggested that ACRE plays an important role in the mechanism regulating the sugar-responsible gene expression through the CMSRE-1 conserved across plant species.

In silico design of CRISPR/Cas9 guide RNA for the knockout of the phytoene desaturase gene in sweet potato (Ipomoea batatas L.)

This study aimed to design in silico guide RNA (sgRNA) for CRISPR/Cas9-mediated knockout of the phytoene desaturase (PDS) gene in sweet potato (Ipomoea batatas L.). The sequence of the coding region of the IbPDS gene is 1791 base pairs (bp) long, and these, in turn, are equivalent to 572 amino acids. The amino acid sequence of the IbPDS gene was compared with the homologous sequences of other nearby plant species, showing that it presents a close similarity with PDS of Ipomoea triloba and Ipomoea nil with 98.60% and 97.73%, respectively. CRISPR RGEN Tools provided 113 results for the IbPDS gene, filtering to 24 and selecting three sgRNA sequences for the design of the gene editing vector, which were sgRNA 1 (5'-ACCTCATCAGTCACCCTGTCNGG-3'), sgRNA 2 (5'- CCTCCAGCAGCAGTATTGGTTGGTTTGNGG -3') and sgRNA 3 (5'- CTGAACTCTCCTGGTTGGTTGTTNGG -3'). The predicted secondary structures of the selected sgRNAs present efficient sgRNA structures for gene editing of the target gene. The PMH-Cas9- 3xsgRNA vector for CRISPR/Cas9-mediated knockout of the IbPDS gene was designed in silico with three sgRNA sequences and one Hygromycin resistance marker. Keywords: Gene editing, sgRNA, IbPDS, gene editing vector, Hygromycin.

Labor- and cost-effective long-read amplicon sequencing using a plasmid analysis service: Application to transposon-inserted alleles in Japanese morning glory.

The sequencing of PCR fragments amplified from specific regions of genomes is a fundamental technique in molecular genetics. Sanger sequencing is commonly used for this analysis; however, amplicon sequencing utilizing next-generation sequencing has become widespread. In addition, long-read amplicon sequencing, using Nanopore or PacBio sequencers to analyze long PCR fragments, has emerged, although it is often more expensive than Sanger sequencing. Recently, low-cost commercial services for full-length plasmid DNA sequencing using Nanopore sequencers have been launched in several countries, including Japan. This study explored the potential of these services to sequence long PCR fragments without the need for cloning into plasmid DNA, as cloning long PCR fragments or blunt-end PCR fragments into plasmid DNA is often challenging. PCR fragments of 4-11 kb, amplified from the DFR-B gene involved in the biosynthesis of anthocyanin, with or without Tpn1 transposons in Japanese morning glory (Ipomoea nil), were circularized using T4 DNA ligase and analyzed as templates. Although some inaccuracies in the length of homopolymer stretches were observed, the remaining sequences were obtained without significant errors. This method could potentially reduce the labor and costs associated with cloning, primer synthesis, and sequence assembly, thus making it a viable option for the analysis of long PCR fragment sequences. Moreover, this study reconfirmed that Tpn1 transposons are major mutagens in I. nil and demonstrated their transposition in the Violet line, a long-used standard in plant physiology.

Active defense strategies for invasive plants may alter the distribution pattern of pests in the invaded area.

In the invaded areas, it is believed that invasive species reduce their investment in defense due to the absence of natural enemies. By field investigation and a series of laboratory assays, This study explored the defense strategies of invasive plants. Field investigation indicated that invasive plants have a antifeedant effect on herbivorous pests, and the distribution frequency of wormholes of native plants shows a peak at a distance of 2-3 m from the invasive species. The feeding preference experiment conducted with two generalist herbivorous insects (native insect Spodoptera litura and invasive insect Spodoptera frugiperda) showed that the invasive plants have a stronger antifeedant effect than native plants. By analyzing the content of secondary metabolites in the leaves of three invasive plants (Sphagneticola trilobata, Mikania micrantha, Ipomoea cairica) and three native plants (Ipomoea nil, Paederia foetida, Polygonum chinense), the leaves of invasive plants had higher concentrations of substances associated with defenses, including total phenols, flavonoids, jasmonic acid, tannin, H2O2, and total antioxidant capacity (TAC), and lower soluble protein concentrations than native plants. After leaf damage, compared to native plants, the leaves of invasive plants showed an overall increase in substances associated with defense, except for soluble sugar. These results suggest that invasive plants maintain active defense strategies in invaded areas, leading to changes in the distribution patterns of herbivorous insects in a manner that facilitates invasion.

Expression Analysis and Interaction Protein Screening of CoZTL in Camellia oleifera Abel

Camellia oleifera Abel., which produces fruits of high comprehensive utilization value, is an important woody oil tree in China. ZEITLUPE (ZTL) is a blue light receptor and clock component protein that is involved in various physiological and biochemical processes. However, the expression pattern and function of C. oleifera ZTL (CoZTL) remain unclear. In this study, the coding sequence of the CoZTL gene was isolated and the protein function was explored using bioinformatics and expression analyses and heterologous expression techniques. The results showed that the CoZTL protein was highly conserved during evolution and was on the same branch of the evolutionary tree as the ZTL proteins from Ipomoea nil and Nicotiana attenuata. CoZTL was mainly expressed in the fruit shells and stems of C. oleifera, and its expression level fluctuated greatly during flower bud development. Transgenic CoZTL-overexpressing Arabidopsis plants showed delayed flowering under long-day conditions as well as light-dependent promotion of hypocotyl elongation. Furthermore, yeast two-hybrid library screening revealed that seven C. oleifera proteins (CoAAT, Coβ-GAL, CoLAT52-like, CoCAR4-like, CoAO, CoUQCC1, and CoADF 2) interacted with CoZTL. Our results indicate that CoZTL plays an important role in C. oleifera flowering and hypocotyl growth.

Promotion of droplet deposition, diffusion-wetting and retention on hydrophobic surfaces by nonionic star-shaped oligomeric surfactants

Massive wastage of pesticides has caused significant environmental pollution and economic loss. However, existing strategies to control loss of pesticides are complex or step-specific. Here, nonionic star-shaped surfactants that can simultaneously achieve comprehensive regulation of deposition, diffusion-wetting and retention are designed and synthesized, which greatly improve the utilization efficiency of pesticides while reducing environmental risks. Structure–activity relationships for the surfactants were elucidated, and their interactions with hydrophobic surfaces were studied in detail. The results show that the addition of nonionic star-shaped oligomeric surfactant to a solution decreased droplet bouncing, increased the diffusion-wetting area, slowed droplet evaporation; and the surfactant III-8b showed the best performance. On the basis of these results, III-8b was used as a synergist for Glyphosate-isopropylammonium 41 % AS. The biological activity test showed that the herbicidal activities of the surfactant III-8b-containing glyphosate solution against Pharbitis nil (L.) Choisy and Echinochloa crusgalli (L.) were substantially better than the activities in the absence of surfactant.

Investigation of resistance mechanisms to fomesafen in Ipomoea nil from China

Recently, the herbicide fomesafen has frequently failed to control the troublesome weed Ipomoea nil in soybean fields in Liaoning Province, China. Hence, we collected 10 suspected resistant populations and evaluated their sensitivity to fomesafen. The results revealed various degrees of Ipomoea nil resistance to fomesafen, with a resistance index of 2.88 to 22.43; the highest value occurred in the LN3 population. Therefore, the mechanisms of the resistance in LN3 to fomesafen were explored. After fomesafen treatment, the expression levels of InPPX1 and InPPX2 genes were 4.19- and 9.29-fold higher, respectively, in LN3 than those in the susceptible (LN1) population. However, mutations and copy number variations were not detected between the two populations. Additionally, malathion pretreatment reduced the dose necessary to halve the growth rate of LN3 by 58%. Liquid chromatography with tandem mass spectrometry demonstrated that metabolism of fomesafen was significantly suppressed by malathion. Moreover, LN3 displayed increased reactive oxygen species scavenging capacity, which was represented by higher superoxide dismutase and peroxidase activities after fomesafen application than those in LN1. An orthogonal partial least squares-discriminant analysis revealed that the high resistance in LN3 could be attributed mainly to enhanced metabolism. Fortunately, the fomesafen-resistant I. nil remained sensitive to 2,4-D-ethylhexylester and bentazon, providing methods for its control.

Transcriptional profiling of long noncoding RNAs associated with flower color formation in Ipomoea nil.

LncRNAs regulate flower color formation in Ipomoea nil via vacuolar pH, TCA cycle, and oxidative phosphorylation pathways. The significance of long noncoding RNA (lncRNA) in diverse biological processes is crucial in plant kingdoms. Although study on lncRNAs has been extensive in mammals and model plants, lncRNAs have not been identified in Ipomoea nil (I. nil). In this study, we employed whole transcriptome strand-specific RNA sequencing to identify 11,203 expressed lncRNA candidates, including 961 known lncRNA and 10,242 novel lncRNA in the I. nil genome. These lncRNAs in I. nil had fewer exons and were generally shorter in length compared to mRNA genes. Totally, 1141 different expression lncRNAs (DELs) were significantly identified between white and red flowers. The functional analysis indicated that lncRNA-targeted genes were enriched in the TCA cycle, photosynthesis, and oxidative phosphorylation-related pathway, which was also found in differentially expressed genes (DEGs) functional enrichments. LncRNAs can regulate transcriptional levels through cis- or trans-acting mechanisms. LncRNA cis-targeted genes were significantly enriched in potassium and lysosome. For trans-lncRNA, two energy metabolism pathways, TCA cycles and oxidative phosphorylation, were identified from positive association pairs of trans-lncRNA and mRNA. This research advances our understanding of lncRNAs and their role in flower color development, providing valuable insights for future selective breeding of I. nil.

First Report of Leaf Spot on Ipomoea nil Caused by Alternaria alternata in China.

Ipomoea nil (Linnaeus) Roth, belonging to the Convolvulaceae family, is an ornamental and medicinal plant in China, which has the function of diuretic and expectorant, and it is also a common weed in the field. In October 2021, a leaf spot disease was observed on I. nil in a field as weed in Jingzhou (N 30° 21', E 112° 19'), Hubei Province, China. Symptoms began as small brown blotches, then developed into oval or irregularly shaped brown necrotic lesions. In severe cases, the leaves were completely necrotic and detached. In the surveyed area, the incidence was between 30% - 40%. To isolate the pathogen, twenty-one leaf pieces (5×5 mm) were cut from the lesion edges of seven symptomatic leaves, disinfected with 70% ethanol and 2% sodium hypochlorite (NaOCl), rinsed with sterile water five times, then placed on three potato dextrose agar (PDA) modified with 50 μg/mL kanamycin, and incubated at 25 °C in dark for 5 days. The isolates were subcultured by transferring mycelium tips. Sixteen fungal strains were isolated from the tissues, and nine of them showed similar morphological characteristics. After cultured 7 days on PDA at 25 °C, the nine colonies were initially white, then turned greenish brown to black in the center and had abundant fine villous aerial mycelia up to 61.5 mm in average diameter. To examine its conidial morphology, the fungi were cultured for 7 days on potato carrot agar (PCA) at 22°C with a light/dark period of 8/16 h. On PCA, conidia were brown or olive-brown, obclavate to obpyriform, with a short beak, one to five transverse and zero to three longitudinal septa. They formed chains of 1 - 8 conidia, with branches. Conidia were 16 - 46 µm long and 8 - 14 µm wide (n=50). These morphological features were similar to those described in Alternaria spp. (Simmons 2007). A single isolate "Q2" was selected for molecular identification because it was the most aggressive in preliminary leaf pathogenicity assays. The internal transcribed spacer (ITS) region of rDNA and histone 3 (H3) gene were amplified and sequenced using primers ITS1/ITS4 (White et al. 1990) and H3-1a/H3-1b (Zheng et al. 2015). BLAST analysis revealed that the sequences (ITS, ON360984; H3, ON375577) were 100% identical to Alternaria alternata (ITS, MK396607; H3, MN840996), respectively. Maximum likelihood analysis based on combined two gene sequences was conducted with an evolutionary model of GTR+I+G under 1000 bootstrap replicates. Phylogenetic tree showed that Q2 and Alternaria alternata 21-5 and BLH-YB-11 located in one clade supported with 99% bootstrap values. The pathogen was identified as A. alternata. To fulfill Koch's postulate, 10 ml conidia (106 spores/ml) of Q2 was sprayed on five healthy seedlings, with sterile distilled water as a control. All leaves were rinsed three times with sterile water before inoculation. All seedlings were placed in sealed plastic bags with air valves, and grown in a greenhouse (25 ± 2 ˚C, RH 65%). The test was repeated twice. After 10 days, symptoms typical of brown blotches similar to those observed in the field were observed on leaves of inoculated plants, while control remained healthy. A. alternata was re-isolated from the inoculated symptomatic leaves with a frequency of 100% based on morphological and molecular characters, thus Koch's postulate was confirmed. To the best of our knowledge, this is the first report of A. alternata causing leaf spot on I. nil in China. Our findings extended the host range of the pathogen A. alternata on characteristic plants.

Genome-Wide Analysis of Aquaporins in Japanese Morning Glory (Ipomoea nil)

The aquaporin (AQP) family, also called water channels or major intrinsic proteins, facilitate water transport. AQPs also transport low-molecular-weight solutes, including boric acid, glycerol, urea, and ammonia. Since plants are sessile, water homeostasis is crucial. Therefore, plants have developed diverse AQP variants at higher expression levels than animals. For example, 35 and 33 AQPs have been identified in Arabidopsis and rice, respectively. In the present study, we identified AQPs in morning glory (Ipomoea nil), which has been widely used as a model plant in research on flowering and floral morphology. The importance of AQPs in the opening of morning glory flowers has been reported. In the morning glory genome, 44 AQPs were identified, and their characteristics were analyzed. A phylogenetic analysis revealed five AQP subfamilies in morning glory: plasma membrane-intrinsic proteins (PIPs), tonoplast-intrinsic proteins (TIPs), nodulin 26-like intrinsic proteins (NIPs), small basic intrinsic proteins (SIPs), and X-intrinsic proteins (XIPs). Further, transport substrates of morning glory AQPs were estimated based on their homology to the known AQPs in other plant species and their corresponding amino acid motifs that possess permeability pores. It was expected that PIPs are likely to transport water, carbon dioxide, and hydrogen peroxide; TIPs are likely transport water, hydrogen peroxide, ammonia, urea, and boric acid; NIPs are likely transport water, boric acid, ammonia, glycerol, and formamide; and XIPs are likely to transport water, hydrogen peroxide, and glycerol. Overall, these results suggest that AQPs are involved in water and nutrient transport in Japanese morning glory. An in silico gene expression analysis suggested the importance of AQPs in flower opening, water or nutrient uptakes from the soil to roots, and photosynthesis in morning glory. Our findings provide fundamental information that enables further study into the importance of AQPs in morning glory, including their roles in flower opening and other physiological events.

Discrimination of morningglory species (Ipomoea spp.) using near-infrared spectroscopy and multivariate analysis

AbstractThe occurrence of weeds is one of the main factors limiting agricultural productivity. Studies on new techniques for the identification of these species can contribute to the development of proximal sensors, which in the future might be coupled to machines to optimize the performance of species-specific weed management. Thus, the objective of this study was to use near-infrared (NIR) spectroscopy and multivariate analysis to discriminate three morningglory species (Ipomoea spp.). The NIR spectra were collected from the leaves of the three weed species at the vegetative stage (up to five leaves), within the spectral band of 4,000 to 10,000 cm−1. The discrimination models were selected according to accuracy, sensitivity, specificity, and Youden’s index and were analyzed with a validation data set (n = 135). The best results occurred when the selection of spectral bands associated with the use of preprocessing was performed. It was possible to obtain an accuracy of 99.3%, 98.5%, and 98.7% for ivyleaf morningglory (Ipomoea hederifolia L.), Japanese morningglory [Ipomoea nil (L.) Roth], and hairy woodrose [Merremia aegyptia (L.) Urb.], respectively. NIR spectroscopy associated with principal component analysis and linear discriminant analysis (PC-LDA) or partial least-squares regression with discriminant analysis (PLS-DA) can be used to discriminate Ipomoea spp.

Bioassay-Guided Isolation of Anthelmintic Components from Semen pharbitidis, and the Mechanism of Action of Pharbitin.

Parasitic helminths continue to pose problems in human and veterinary medicine, as well as in agriculture. Semen pharbitidis, the seeds of Pharbitis nil (Linn.) Choisy (Convolvulaceae), is a well-known traditional Chinese medicinal botanical preparation widely used for treating intestinal parasites in China owing to its desirable efficacy. However, the anthelmintic compounds in Semen pharbitidis and their mechanism of action have not been investigated yet. This study aimed to identify the compounds active against helminths from Semen pharbitidis, and to establish the mechanism of action of these active compounds. Bioassay-guided fractionation was used to identify the anthelmintic compounds from Semen pharbitidis. The anthelmintic assay was performed by monitoring Caenorhabditis elegans (C. elegans) motility with a WMicrotracker instrument. Active compounds were identified by high-resolution mass spectrometry. Several (analogues of) fragments of the anthelmintic compounds were purchased and tested to explore the structure-activity relationship, and to find more potent compounds. A panel of C. elegans mutant strains resistant to major currently used anthelmintic drugs was used to explore the mechanism of action of the active compounds. The bioassay-guided isolation from an ethanol extract of Semen pharbitidis led to a group of glycosides, namely pharbitin (IC50: 41.0 ± 9.4 μg/mL). Hit expansion for pharbitin fragments yielded two potent analogues: 2-bromohexadecanoic acid (IC50: 1.6 ± 0.7 μM) and myristoleic acid (IC50: 35.2 ± 7.6 μM). One drug-resistant mutant ZZ37 unc-63 (x37) demonstrated a ~17-fold increased resistance to pharbitin compared with wild-type worms. Collectively, we provide further experimental scientific evidence to support the traditional use of Semen pharbitidis for the treatment of intestinal parasites. The anthelmintic activity of Semen pharbitidis is due to pharbitin, whose target could be UNC-63 in C. elegans.

Eco-friendly bio-encapsulation from sodium alginate-trehalose-kaolin and its performance evaluation in improving plant growth under salt or/and drought conditions

Plant growth-promoting bacterium plays a significant role in improving plant tolerance to abiotic stresses. However, there are low survival and poor effect in field application, especially in unfavorable environments. Our previous study suggested that encapsulation of Bacillus pumilus G5 from polyvinyl alcohol‑sodium alginate could improve plant growth and soil fertility under drought and salt soil conditions. However, in the G5 microbeads, the polyvinyl alcohol could not be degraded after entering the soil, and the loss of viable bacteria was severe during the drying process. Achieving a more eco-friendly and efficient formulation based on biodegradable polymers can have significant effects on increasing the quantity and quality of agricultural products. Herein, G5 has immobilized in the composite wall of sodium alginate-trehalose-kaolin microbeads and then evaluated the performance, and applied on the Pharbitis nil under salt or/and drought stress by pot experiment. A 2 % sodium alginate, 1 % trehalose, and 1 % kaolin formulation for the coating films resulted in optimal G5 microbeads embedding efficiency, viable bacteria, degradation rate, and sustained release. Also, the G5 microbeads exhibited longer storage life than that of the G5 suspension. Scanning electron microscopy revealed that the G5 microcapsules had a near-spherical structure with a particle size of around 1000 μm forming a continuous dense composite wall membrane with obvious protrusions and folds on the surface, which facilitated the release of the G5 strain. The interior of the G5 capsule was rough and suitable for bacterial attachment. Infrared spectroscopy showed that the G5 microcapsules are a simple physical mixture with no chemical reaction between the excipients, making the G5 microcapsules chemically stable. The inclusion of the G5 microcapsules considerably induced Pharbitis nil seedlings growth and biomass under drought and/or salt stress. In the rhizosphere soil of Pharbitis nil, the G5 microcapsules increased the total cultivable bacteria population, the activities of invertase, urease, phosphatase, and catalase, and the contents of available nitrogen and available phosphorus. We concluded that a suitable formulation by bio-encapsulation with eco-friendly excipients for alleviating drought and/or salt stress in plants will be advantageous in sustainable agriculture.

Liana and vine germination requirements in a subtropical forest zone

Climbing plants may be classified as lianas (woody) or vines (herbaceous). This grouping is not taxonomic but is useful for functional ecology. We evaluated seed germination of three liana and four vine species from the Chinese subtropical forest zone. Seeds were collected from different environments (abandoned agricultural land, broadleaf forest, coniferous mixed forest) and seed traits characterized (seed fresh mass, seed shape index, water gain). Germination tests were conducted under different temperature regimes: 5/10, 10/20, 20/30, 25/35, 35/40 °C under light and dark conditions. Additionally, we tested the relationships of germination responses (germination percentage, mean germination times, and relative light-germination index) with seed traits. The two species from abandoned agricultural land (Glycine max subsp. soja and Ipomoea triloba) had impermeable seed coats and reached only intermediate values of germination percentage, from 20 to 40% in the tested temperatures, with little effect from light regimes. In the broadleaf forest, Ipomoea nil also had impermeable seeds, but reached 91% of germination under 25/35 °C. Seeds of I. nil were the largest ones (4.8 g per 100 seeds), and there was a positive relationship of seed mass with G%. Seeds of Kadsura longipedunculata (liana) had low G% under all tested temperatures, probably related to some dormancy type, which requires further investigation. Species from coniferous mixed forest were greatly affected by temperature regimes, and Coptosapelta diffusa (the smallest seeds) had also the most light-dependent germination. Light-germination index had thus a significant/negative relationship with seeds mass. Environmental factors may help to explain the distribution of seed traits in our study, but further research should examine a larger set of species to better understand regeneration ecology of climbing plants in contrasting ecosystems.