Establishment of an Artificial Inoculation Method of Ustilaginoidea virens Without Damaging the Rice Panicle Sheaths.

Rice false smut (RFS) is a destructive disease of rice worldwide caused by Ustilaginoidea virens. Nevertheless, there is a lack of efficient and stable artificial inoculation method to simulate the natural infection of U. virens, which is an important factor limiting further research on the pathogen. The purpose of this study was to establish an artificial inoculation method, which can simulate the natural infection process of U. virens without destroying the panicle sheath structure of rice. In this research, rice plants were inoculated by soaking roots at the seedling stage, spraying at the tillering stage, injecting at the booting stage, and again spraying at the flowering stage to determine the appropriate artificial inoculation time. Meanwhile, the panicle sheath instillation method and the injection inoculation method were compared. The results show that stages 6 to 8 of young panicle differentiation are an important period for U. virens infection. There were no significant differences in the mean rates of infected panicles, mean rates of infected grains, and maximum infected grains per panicle between the two inoculation methods. However, the frequency of RFS ball occurrence at the upper part of the panicles was significantly higher on the spikelets inoculated by the injection method than that of spikelets inoculated by natural infection and panicle sheath instillation. Therefore, panicle sheath instillation method was more similar to the natural infection of U. virens in the field. This research exhibited an innovative artificial inoculation method for identification of U. virens pathogenicity and evaluation of rice resistance against RFS.

Repellency Mechanism of Natural Guar Gum-Based Film Incorporated with Citral against Brown Planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae).

Using of plant essential oil that coevolved as a defense mechanism against agriculture insects is an alternative means of controlling many insect pests. In order to repel brown planthoppers (BPHs), the most notorious rice insect pest, a new film based on guar gum incorporated with citral (GC film) was formulated, which was effective while being environmentally friendly. In this paper, the effect and mechanism of GC film repellency against BPHs were determined. Repellent activity test and olfactory reaction analysis showed that GC film had repellency effect against BPHs, with repellency of 60.00% and 73.93%, respectively. The result of olfactory reaction indicated that GC film repellency against BPHs relied on smell. EPG analysis showed the proportion and mean duration of np waveform were significantly higher than in CK and increased following the treatment concentration, which indicated that GC film affected the recognition of BPHs to rice. Further analysis by RNA sequencing analysis showed a total of 679 genes were significantly upregulated and 284 genes were significantly downregulated in the BPHs fed on the rice sprayed with GC film compared to control. Odorant-binding protein (OBP) gene 797 and gustatory receptor gene (GR)/odorant receptor (OR) gene 13110 showed a significant decrease in differential expression and significant increase in differential expression, respectively. There were 0.66 and 2.55 differential expression multiples between treated BPHs and control, respectively. According to the results described above, we reasoned that GC film repellency against BPHs due to smell, by release of citral, caused the recognition difficulties for BPHs to rice, and OBP gene 797 and GR/OR gene 13110 appeared to be the crucial candidate genes for GC film repellency against BPHs. The present study depicted a clear and consistent repellency effect for GC film against BPHs and preliminarily clarified the mechanism of GC film as a repellent against BPHs, which might offer an alternative approach for control of BPHs in the near future. Our results could also help in the development and improvement of GC films.

Physicochemical Properties and Bioactivity of a New Guar Gum-Based Film Incorporated with Citral to Brown Planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae).

The brown planthopper (BPH), Nilaparvata lugens (Stål), is the most notorious rice insect pest. In order to repel BPH effectively while being environmentally friendly, a new film based on guar gum incorporated with citral (GC film) was formulated. A toxicity bioassay of citral and guar gum at different proportions (ratios of 3:1, 2:1, 1:1, 1:2, and 1:3 in w/w) of GC film-forming emulsion to BPH was performed with the rice stem dipping method. Results showed that the most effective ratio of citral to guar gum was 1:1 with the median lethal concentration (LC50) of 4.30 mg/mL, far below the LC50 of guar gum (GG)/citral individual (141.51 and 44.38 mg/mL, respectively). The mortality of BPH adults and nymphs in the third instar treated with different dilution multiples of GC film-forming emulsion ranged from 46.67% to 82.22% and from 37.78% to 71.11%, respectively. These indicated that GC film-forming emulsion had a direct toxicity on BPH, and the mixture of citral and GG had synergistic interactions. Subsequently, Fourier-transform infrared spectroscopy showed that the incorporation of guar gum with citral was successful and did not result in the formation of new chemical bonds. The GC film exhibited a darker color and rougher surface topography with larger apertures and deeper gullies (Ra = 1.42 nm, Rq = 2.05 nm, and Rmax = 25.40 nm) compared to the guar gum film (GG film) (Ra = 1.00 nm, Rq = 1.33 nm, and Rmax = 16.40 nm), as determined by transmission electron microscopy and atomic force microscopy. The GC film exhibited a 50.4% lower solubility in water (30.30% vs. 15.00%) and 71.3% oxygen permeability (8.26 × 10-9 vs. 2.37 × 10-9 cm3/m2·d·Pa) (p < 0.05) but did not demonstrate any significant difference in mechanical properties, such as thickness (39.10 vs. 41.70 mm), tensile strength (41.89 vs. 38.30 N/mm2), and elongation at break (1.82% vs. 2.03%) (p < 0.05) compared to the GG film. Our findings established a link between physicochemical properties and bioactivity, which can provide useful information on developing and improving GC films and may offer an alternative approach for the control of BPH in the near future.

Effects of introducing theanine or glutamic acid core to tralopyril on systemicity and insecticidal activity.

Tralopyril was the active agent of a pro-insecticide chlorfenapyr. To simultaneously solve the problems of the phytotoxicity and non-systemic insecticidal activity of tralopyril, four new tralopyril conjugates containing theanine or glutamic acid moieties were designed and synthesized. Their phytotoxicity to tea shoot, phloem systemicity, and insecticidal activity were evaluated. Phytotoxic symptoms were not observed after the tea shoots were exposed to the four conjugates at concentrations of 2mM. The phloem mobility test on Ricinus communis L. seedlings confirmed that all four conjugates were mobile in the sieve tubes. Results of insecticidal activity against the third-instar larvae of Plutella xylostella showed that only conjugate 20 exhibited activity with an LC50 value of 0.5882±0.0504mM. After root application to tea seedlings, conjugate 20 showed obviously systemic insecticidal activity against Dendrothrips minowai Priesner, while chlorfenapyr showed no attribute of that. A new conjugate as potential phloem mobile pro-insecticide candidate was provided and so a novel strategy of pro-insecticide for improved phloem systemicity was proposed.

Individual and combined effects of arbuscular mycorrhizal fungi and phytohormones on the growth and physiobiochemical characteristics of tea cutting seedlings.

Both arbuscular mycorrhizal fungi (AMF) and phytohormones collectively regulate plant growth and root development, but their individual and combined effects on tea [Camellia sinensis (L.) O. Kuntze] cutting seedings remain unclear. This study examined the individual and combined effects of two species of AMF (Rhizophagus intraradices, RI and Funneliformis mosseae, FM) and two types of palnt hormones (strigolactones, SLs; polyamines, PAs) on tea cutting seedings, by evaluating the growth and physiobiochemical characteristics of plants treated with the AMFs and/or hormones. The results showed that inoculation with either AMF individually or hormones treatment alone could significantly enhanced mycorrhizal colonization, growth target and physiobiochemical characteristics of tea cutting seedlings. Interestingly, the addition of a combination of AMFs and hormones showed superior effects, while SL and RI exhibited the most improvements to the colonization rate, plant growth, root-morphological traits, root DHA activity, photosynthesis, chlorophyll content, soluble sugar content in leaves, and the activities of antioxidant enzymes (SOD, POD, and CAT), compared to other treatment combinations (SL + FM, PA + RI, and PA + FM). Correlation analyses revealed a significantly (p < 0.05) positive correlation of root AMF colonization with root-related traits (e.g., DHA, root total length, surface area, and volume) and leaf-related traits (e.g., leaf area, shoot biomass, total chlorophyll, and antioxidant enzyme activities). This study demonstrated that while the apllication of individual AMF or plant hormones had a certain good effects on most growth and physiobiochemical characteristics parameters of tea cutting seedings, the additive effect was from specific combined of AMF and plant hormones. These results highlight the possibility for combined of AMF and plant hormones to improve the asexual reproduction of tea plants via cuttings.