Metabolic pathways modulated by coumarin to inhibit seed germination and early seedling growth in Eleusine indica.

Coumarin is an allelochemical that is widely present in the plant kingdom and has great potential for weed control. However, its mechanisms of action remain largely unknown. This study employed metabolomic and transcriptomic analyses along with evaluations of amino acid profiles and related physiological indicators to investigate how coumarin inhibits the germination and seedling growth of Eleusine indica by modifying metabolic pathways. At 72 h of germination at 50 and 100 mg L-1 coumarin, E. indica had lower levels of soluble sugar and activities of amylases and higher levels of starch, O2-, H2O2, auxin (IAA) and abscisic acid (ABA) compared to the control. Metabolomic analysis demonstrated that coumarin treatments had a significant impact on the pathways associated with amino acid metabolism and transport and aminoacyl-tRNA biosynthesis. Exposure to coumarin induced significant alterations in the levels of 19 amino acids, with a decrease in 15 of them, including Met, Leu and γ-aminobutyric acid (GABA). Additionally, transcriptomic analysis showed that coumarin significantly disrupted several essential biological processes, including protein translation, secondary metabolite synthesis, and hormone signal transduction. The decrease in TCA cycle metabolite (cis-aconitate, 2-oxoglutarate, and malate) contents was associated with the suppression of transcription for related enzymes. Our findings indicate that the inhibition of germination and growth in E. indica by coumarin involves the suppression of starch conversion to sugars, modification of the amino acid profile, interference of hormone signalling and the induction of oxidative stress. The TCA cycle appears to be one of the most essential pathways affected by coumarin.

Temperature influences glyphosate efficacy on glyphosate-resistant and -susceptible goosegrass (Eleusine indica).

Glyphosate has been widely used to control Eleusine indica and other weeds in South China for many years. Among the most troublesome weeds in South China, E. indica can remain alive all year round. However, the influence of temperature on glyphosate efficacy on E. indica, especially under days with fluctuating temperature, is unknown. This study evaluated the influence of two temperature regimes on glyphosate efficacy on glyphosate-resistant (R) and -susceptible (S) E. indica biotypes. Plants of the R and S biotypes were cultivated under two temperature regimes (high: 30°C/20°C day/night; low: 20°C/15°C day/night). Dose-response experiments showed improved efficacy of glyphosate at the low temperature compared with that at the high temperature for both biotypes. Based on the LD50 values, the R biotype was 8.9 times more resistant to glyphosate than the S biotype at the high temperature; however, the resistance index (R/S) decreased to 3.1 at the low temperature. At 4 days after glyphosate application, shikimic acid accumulation was greater at the low temperature than at the high temperature in plants of both biotypes, and the increase was higher in plants of the R biotype than in the S biotype. At a sublethal glyphosate dose (R: 400 g ai ha-1; S: 200 g ai ha-1), plants grown at the low temperature showed a strong decrease in leaf chlorophyll content and Fv/Fm value compared with those of plants grown at the high temperature and the untreated control. At 3 days after treatment, glyphosate absorption was similar between biotypes at the high temperature, but absorption decreased to 64.9% and 53.1% at the low temperature for the R and S biotypes, respectively. For both biotypes, glyphosate translocation from the leaf to the remainder of the plant was reduced at the low temperature compared with that at the high temperature. No differences in glyphosate translocation were observed between biotypes within each temperature regime. This is the first report on the effect of temperature on glyphosate efficacy on E. indica, and provides important insights for glyphosate application and resistance management.

Diel dynamics of multi-omics in elkhorn fern provide new insights into weak CAM photosynthesis.

Crassulacean acid metabolism (CAM) has high water-use efficiency (WUE) and is widely recognized to have evolved from C3 photosynthesis. Different plant lineages have convergently evolved CAM, but the molecular mechanism that underlies C3-to-CAM evolution remains to be clarified. Platycerium bifurcatum (elkhorn fern) provides an opportunity to study the molecular changes underlying the transition from C3 to CAM photosynthesis because both modes of photosynthesis occur in this species, with sporotrophophyll leaves (SLs) and cover leaves (CLs) performing C3 and weak CAM photosynthesis, respectively. Here, we report that the physiological and biochemical attributes of CAM in weak CAM-performing CLs differed from those in strong CAM species. We investigated the diel dynamics of the metabolome, proteome, and transcriptome in these dimorphic leaves within the same genetic background and under identical environmental conditions. We found that multi-omic diel dynamics in P. bifurcatum exhibit both tissue and diel effects. Our analysis revealed temporal rewiring of biochemistry relevant to the energy-producing pathway (TCA cycle), CAM pathway, and stomatal movement in CLs compared with SLs. We also confirmed that PHOSPHOENOLPYRUVATE CARBOXYLASE KINASE (PPCK) exhibits convergence in gene expression among highly divergent CAM lineages. Gene regulatory network analysis identified candidate transcription factors regulating the CAM pathway and stomatal movement. Taken together, our results provide new insights into weak CAM photosynthesis and new avenues for CAM bioengineering.

Evolution of multiple target-site resistance mechanisms in individual plants of glyphosate-resistant Eleusine indica from China.

BACKGROUND: Glyphosate has been used for weed control in South China in various situations for four decades, and most Eleusine indica populations are suspected to have evolved resistance to glyphosate. This research investigated underling target-site glyphosate resistance mechanisms in six field-collected, putative glyphosate-resistant (R) E. indica populations. RESULTS: The six R E. indica populations were confirmed to be low (1.8 to 2.6-fold) to moderately (5.6- to 8.4-fold) resistant to glyphosate relative to the susceptible (S) population. Sixty-seven glyphosate-surviving plants from the six R populations were used to examine target-site resistance mechanisms. Target-site 5-enolpyruvylshikimate3-phosphate synthase (EPSPS) overexpression (OE) (plus further induction by glyphosate treatment) and gene copy number variation (CNV) occurred in 94% R plants, and among them, 16% had the P106A mutation and 49% had the heterozygous double TIPS (T102I + P106S) mutation (plus P381L). In addition, a low number of R plants (6%) only had the homologous TIPS (plus P381L) mutation. The (CT)6 insertion mutation in the EPSPS 5†-UTR always associates with EPSPS OE and CNV. Progeny plants possessing EPSPS OE/CNV (and P106A) displayed low level (up to 4.5-fold) glyphosate resistance. In contrast, plants homozygous for the TIPS mutation displayed higher (25-fold) resistance to glyphosate and followed by plants heterozygous for this mutation plus EPSPS OE/CNV (12-fold). CONCLUSIONS: Target-site glyphosate resistance in E. indica populations from South China is common with prevalence of EPSPS OE/induction/CNV conferring low level resistance. Individual plants acquiring both the TIPS mutation and EPSPS OE/CNV are favored due to evolutionary advantages. The role of (CT)6 insertion mutation in EPSPS CNV is worth further investigation. © 2021 Society of Chemical Industry.

Strong Response of Stem Photosynthesis to Defoliation in Mikania micrantha Highlights the Contribution of Phenotypic Plasticity to Plant Invasiveness.

Phenotypic plasticity affords invasive plant species the ability to colonize a wide range of habitats, but physiological plasticity of their stems is seldom recognized. Investigation of the stem plasticity of invasive plant species could lead to a better understanding of their invasiveness. We performed pot experiments involving defoliation treatments and isolated culture experiments to determine whether the invasive species Mikania micrantha exhibits greater plasticity in the stems than do three non-invasive species that co-occur in southern China and then explored the mechanism underlying the modification of its stem photosynthesis. Our results showed that the stems of M. micrantha exhibited higher plasticity in terms of either net or gross photosynthetic rate in response to the defoliation treatment. These effects were positively related to an increased stem elongation rate. The enhancement of stem photosynthesis in M. micrantha resulted from the comprehensive action involving increases in the Chl a/b ratio, D1 protein and stomatal aperture, changes in chloroplast morphology and a decrease in anthocyanins. Increased plasticity of stem photosynthesis may improve the survival of M. micrantha under harsh conditions and allow it to rapidly recover from defoliation injuries. Our results highlight that phenotypic plasticity promotes the invasion success of alien plant invaders.

Interpretation of the difference in shade tolerance of two subtropical forest tree species of different successional stages at the transcriptome and physiological levels.

Differences in plant shade tolerance constitute a major mechanism driving the succession of forest communities in subtropical forests. However, the indirect effects of differences in light requirements on the growth of mid- and late-successional tree species are unclear, and this potential growth effect has not been explained at the transcriptome level. Here, a typical mid-successional dominant tree species, Schima superba Gardn. et Champ, and a typical late-successional dominant tree species, Cryptocarya concinna Hance were used as materials and planted under 100% full light (FL) and 30% FL (low light, LL) to explore the responses of tree species in different successional stages of subtropical forests to different light environments. Transcriptome sequencing was used to analyze the expression changes in genes related to growth and photoprotection under different light environments. The young leaves of S. superba accumulated more malondialdehyde (MDA) and superoxide radicals (${\mathrm{O}}_2^{{{}^{\bullet}}^{-}}$) under LL. A lower hormone content (auxin, cytokinin, gibberellin) in the young leaves, a weaker photosynthetic capacity in the mature leaves and significant downregulation of related gene expression were also found under LL, which resulted in the total biomass of S. superba under LL being lower than that under FL. The young leaves of C. concinna had less MDA and ${\mathrm{O}}_2^{{{}^{\bullet}}^{-}}$, and a higher hormone contents under LL than those under FL. There was no significant difference in photosynthetic capacity between mature leaves in contrasting light environments. Although the biomass of C. concinna under LL was less than that under FL, the height of C. concinna under LL was higher than that under FL, indicating that C. concinna could grow well under the two light environments. Our results describing the acclimatization of light at the physiological, molecular and transcriptome levels are important for a complete understanding of successional mechanisms.

[Allelopathic interactions between Borreria latifolia and two common Asteraceae species]. / 阔叶丰花草与2种菊科植物之间的化感作用.

Borrelia latifolia is an annual herb suitable to be used as cover crop for weed control in orchards in southern China. To understand the competition between B. latifolia and common weeds in orchards, we investigated the allelopathic interactions between B. latifolia and two Asteraceae species, Ageratum conyzoides and Bidens alba. The results showed that the aqueous extracts of B. latifolia at 10-50 mg·mL-1 significantly inhibited seed germination and seedling growth of A. conyzoides and B. alba, with the radicle length of A. conyzoides and B. alba being reduced by 57.4%-90.2% and 57.3%-62.3%, respectively. The aqueous extracts of A. conyzoides and B. alba also had strong allelopathic effects on seed germination of B. latifolia. Under the treatment of 50 mg·mL-1 aqueous extracts, the two Asteraceae species almost entirely inhibited the germination of B. latifolia. After treated with 10 mg·mL-1 aqueous extract of B. latifolia for 30 d, net photosynthesis, plant height and biomass of A. conyzoide were reduced by 15.2%, 20.6% and 41.5%, respectively, compared with the control, while the biomass of B. alba also showed a decreasing trend. Instead, the growth of B. latifolia was not affected. Under mixed culture, biomass of B. latifolia was comparable to that under monoculture, whereas those of A. conyzoides and B. alba were reduced by 86.0% and 27.1%, respectively. Compared with A. conyzoides and B. alba, the allelopathic advantage of B. latifolia is that it can inhibit growth of the two Asteraceae species as well as inhibit seed germination.

Seasonal variations in group leaf characteristics in species with red young leaves.

The leaves of many plants are red during particular stages of their lives, but the adaptive significance of leaf colouration is not yet clearly understood. In order to reveal whether anthocyanins play a similar role (i.e. antioxidants) in different seasonal contexts, this study investigated species with red young leaves in the subtropical forest of Dinghushan biosphere reserve (South China) during summer and winter and compared group leaf characteristics between the two seasons. Of 62 total species, 33 exhibited red young leaves in summer only, 6 in winter only, and 23 in both seasons. The anthocyanins extracted from most of these species had an absorption peak at ~530 nm. Frequency distribution analysis showed that the species containing anthocyanins at levels ranging from 0.02 to 0.04 µmol cm-2 occurred most frequently in summer or winter. Based on conditional grouping of the species, no significant variation was observed in the average anthocyanin contents and antioxidant abilities between summer and winter; the flavonoid content in summer was 2-fold that in winter, whereas the anthocyanin:flavonoid ratio in summer was only half that in winter. Moreover, a positive correlation between anthocyanins and flavonoids was found in summer. Therefore, it is less likely for anthocyanins to serve as antioxidants in summer than winter, because such a function in summer leaves is readily replaced by other flavonoids.

Phytotoxic effects of Cu, Cd and Zn on the seagrass Thalassia hemprichii and metal accumulation in plants growing in Xincun Bay, Hainan, China.

Seagrasses play an important role in coastal marine ecosystems, but they have been increasingly threatened by human activities. In recent years, seagrass communities have rapidly degenerated in the coastal marine ecosystems of China. To identify the reasons for the decline in seagrasses, the phytotoxic effects of trace metals (Cu, Cd and Zn) on the seagrass Thalassia hemprichii were investigated, and the environmental contents of the metals were analyzed where the seagrass grows. The results showed that leaf necrosis in T. hemprichii exposed to 0.01-0.1 mg L-1 of Cu2+ for 5 days was more serious than that in plants exposed to the same concentrations of Cd2+ and Zn2+. The chlorophyll content in T. hemprichii declined in a concentration-dependent manner after 5 days of exposure to Cu2+, Cd2+ and Zn2+. The evident reduction in ΔF/Fm' in T. hemprichii leaves was observed at day 1 of exposure to 0.01-1.0 mg L-1 of Cu2+ and at day 3 of exposure to 0.1-1.0 mg L-1 of Cd2+. The antioxidant enzyme activities (SOD, POD and CAT) in T. hemprichii leaves exposed to the three metal ions also showed significant changes. In seawater from Xincun Bay (Hainan, China), where T. hemprichii grows, Cu had reached a concentration (i.e., 0.01 mg L-1) that could significantly reduce chlorophyll content and ΔF/Fm' in T. hemprichii leaves. Our results indicate that Cu influences the deterioration of seagrasses in Xincun Bay.

Functional characteristics of phenolic compounds accumulated in young leaves of two subtropical forest tree species of different successional stages.

The abundance of phenolic compounds (including anthocyanins) in leaves is associated with photosynthetic performance, but the regulatory mechanism is unclear. Schima superba Gardn. et Champ. and Cryptocarya concinna Hance., which exhibit distinct anthocyanin accumulation patterns, are dominant tree species in the early- and late-successional stages, respectively, of subtropical forests in China. RNA-seq and analyses of phenolic concentrations, antioxidant capacity and photosynthetic characteristics were performed on young and mature leaves of these two species under contrasting light conditions. The high-light-acclimated young leaves of S. superba and C. concinna and low-light-acclimated young leaves of C. concinna were red. These red leaves had higher ratios of electron transport rate to gross photosynthesis (ETR:Pgross) and total antioxidant capacity to chlorophyll (TAC:Chl) than did the green leaves, regardless of light conditions. In addition, the red leaves had a higher expression level of the UDP-glucose:flavonoid 3-O-glucosyltransferase (UFGT) gene than did the green leaves, irrespective of light conditions. Total antioxidant capacity was positively correlated with flavonoid content in C. concinna leaves and with total phenolic content in leaves of both species under both high and low light. Consistent with the measurements of photosynthetic performance and flavonoids:Chl ratio, photosynthesis-related genes were extensively downregulated and flavonoid-pathway-related genes were extensively upregulated in young leaves relative to mature leaves. Under high and low light, both non-photochemical quenching and TAC:Chl, which serve as different types of photoprotective tools, were enhanced in young leaves of S. superba, whereas only TAC:Chl was enhanced in young leaves of C. concinna. Our results indicate that the biosynthesis of phenolic compounds in young leaves is likely enhanced by an imbalance between photosynthetic electron supply and demand and that flavonoids play a larger role in meditating photoprotection in late-successional species than in early-successional ones.

A magic red coat on the surface of young leaves: anthocyanins distributed in trichome layer protect Castanopsis fissa leaves from photoinhibition.

The presence of anthocyanins in young leaves plays an important role in mitigation against photodamage and allows leaves to grow and develop normally. Many studies have reported that foliar anthocyanins are distributed within the vacuoles of mesophyll cells, so we explored the novel defence style of anthocyanin-coated young leaves of Castanopsis fissa, a dominant subtropical forest tree species, via removable trichomes. Anthocyanins were distributed in C. fissa leaf trichomes, which produced a red coating for the young leaves. As young leaves developed and then matured, the thickness and density of the anthocyanin trichomes progressively decreased, the coating finally disappearing, allowing greater utilization of light by mature leaves. In addition to anthocyanins, the trichomes contained a remarkably high amount of phenolics, which enable the red coating to be more efficient in screening ultraviolet light. Compared with mature leaves, the young leaves exhibited lower photosynthetic ability, which was attributable to the reduced chlorophyll and Rubisco contents. Removal of the red coating had little effect on the photosynthetic capacity of young leaves. However, the young leaves without the coating suffered greater light-induced photoinhibition due to greater excess light entering the chloroplast and the production of H2O2 Our results suggest that the anthocyanin coating is photoprotective and this anthocyanin defence style may be a metabolically cost-effective way of adjusting the anthocyanin content in response to demand.

A Putative Chloroplast-Localized Ca(2+)/H(+) Antiporter CCHA1 Is Involved in Calcium and pH Homeostasis and Required for PSII Function in Arabidopsis.

Calcium is important for chloroplast, not only in its photosynthetic but also nonphotosynthetic functions. Multiple Ca(2+)/H(+) transporters and channels have been described and studied in the plasma membrane and organelle membranes of plant cells; however, the molecular identity and physiological roles of chloroplast Ca(2+)/H(+) antiporters have remained unknown. Here we report the identification and characterization of a member of the UPF0016 family, CCHA1 (a chloroplast-localized potential Ca(2+)/H(+) antiporter), in Arabidopsis thaliana. We observed that the ccha1 mutant plants developed pale green leaves and showed severely stunted growth along with impaired photosystem II (PSII) function. CCHA1 localizes to the chloroplasts, and the levels of the PSII core subunits and the oxygen-evolving complex were significantly decreased in the ccha1 mutants compared with the wild type. In high Ca(2+) concentrations, Arabidopsis CCHA1 partially rescued the growth defect of yeast gdt1Δ null mutant, which is defective in a Ca(2+)/H(+) antiporter. The ccha1 mutant plants also showed significant sensitivity to high concentrations of CaCl2 and MnCl2, as well as variation in pH. Taken these results together, we propose that CCHA1 might encode a putative chloroplast-localized Ca(2+)/H(+) antiporter with critical functions in the regulation of PSII and in chloroplast Ca(2+) and pH homeostasis in Arabidopsis.

A novel role of ethephon in controlling the noxious weed Ipomoea cairica (Linn.) Sweet.

Several auxin herbicides, such as 2, 4-D and dicamba, have been used to eradicate an exotic invasive weed Ipomoea cairica in subtropical China, but restraining the re-explosion of this weed is still a challenge. Since ethylene is one of the major intermediate functioning products during the eradication process, we explored the possibility, mechanism and efficiency of using ethephon which can release ethylene to control Ipomoea cairica. The results of the pot experiment showed that 7.2 g /L ethephon could totally kill Ipomoea cairica including the stems and roots. The water culture experiment indicated that ethephon released an abundance of ethylene directly in leaves and caused increases in electrolyte leakage, 1-aminocyclopropane-1-carboxylic acid (ACC), abscisic acid (ABA) and H2O2 and decreases in chlorophyll content and photosynthetic activity, finally leading to the death of Ipomoea cairica. The field experiment showed that the theoretical effective concentration of ethephon for controlling Ipomoea cairica (weed control efficacy, WCE = 98%) was 4.06 g/L and the half inhibitory concentration (I50) was 0.56 g/L. More than 50% of the accompanying species were insensitive to the phytotoxicity of ethephon. Therefore, ethephon is an excellent alternative herbicide for controlling Ipomoea cairica.

A new strategy for controlling invasive weeds: selecting valuable native plants to defeat them.

To explore replacement control of the invasive weed Ipomoea cairica, we studied the competitive effects of two valuable natives, Pueraria lobata and Paederia scandens, on growth and photosynthetic characteristics of I. cairica, in pot and field experiments. When I. cairica was planted in pots with P. lobata or P. scandens, its total biomass decreased by 68.7% and 45.8%, and its stem length by 33.3% and 34.1%, respectively. The two natives depressed growth of the weed by their strong effects on its photosynthetic characteristics, including suppression of leaf biomass and the abundance of the CO2-fixing enzyme RUBISCO. The field experiment demonstrated that sowing seeds of P. lobata or P. scandens in plots where the weed had been largely cleared produced 11.8-fold or 2.5-fold as much leaf biomass of the two natives, respectively, as the weed. Replacement control by valuable native species is potentially a feasible and sustainable means of suppressing I. cairica.

Photosynthetic characteristics and light energy conversions under different light environments in five tree species occupying dominant status at different stages of subtropical forest succession.

In order to reveal the mechanism of succession in subtropical forest along a light gradient, we investigated photosynthetic physiological responses to three light environments in five tree species including a pioneer species Pinus massoniana Lamb., two mid-successional species Schima superba Gardn. et Champ. and Castanopsis fissa (Champ. ex Benth.) Rehd. et Wils., and two late-successional species Cryptocarya concinna Hance. and Acmena acuminatissima (BI.) Merr et Perry) that were selected from Dinghu Mountain subtropical forest, South China. Results showed that, among the three kinds of species in all light conditions (100%, 30% and 12% of full sunlight), the pioneer species had the highest photosynthetic capacity (Amax), light saturation point (LSP), carboxylation efficiency (CE) and maximum utilisation rate of triose phosphate (TPU) that characterised a strong photosynthetic capacity and high carbon dioxide uptake efficiency. However, a higher light compensation point (LCP) and dark respiration (Rd) as well as lower apparent quantum yield (AQY) indicated that the pioneer specie cannot adapt to low light conditions. Mid-successional species had photosynthetic characteristics in between pioneer and late-successional species, but had the greatest effective quantum yield of PSII (ΦPSII) and light use efficiency (LUE, expressed in terms of photosynthesis). In contrast to pioneer and mid-successional species, late-successional species had lower photosynthetic capacity and carbon uptake efficiency, but higher shade tolerance and high-light heat dissipation capacity, as characterised by higher levels of total xanthophyll cycle pigments (VAZ) and de-epoxidation state of xanthophyll cycle (DEPs). These results indicate that photosynthetic capacity decreases along the successional axis and that late-successional species have more responsive heat dissipation capability to compensate for their inferior photosynthetic capacity.