Nocturnal burst emissions of germacrene D from the open disk florets of pyrethrum flowers induce moths to oviposit on a nonhost and improve pollination success.

Recent studies show that nocturnal pollinators may be more important to ecosystem function and food production than is currently appreciated. Here, we describe an agricultural field study of pyrethrum (Tanacetum cinerariifolium) flower pollination. Pyrethrum is genetically self-incompatible and thus is reliant on pollinators for seed set. Our pollinator exclusion experiment showed that nocturnal insects, particularly moths, significantly contribute to seed set and quality. We discovered that the most abundant floral volatile, the sesquiterpene (-)-germacrene D (GD), is key in attracting the noctuid moths Peridroma saucia and Helicoverpa armigera. Germacrene D synthase (GDS) gene expression regulates the specific GD production and accumulation in flowers, which, in contrast to related species, lose the habit of closing at night. We did observe that female moths also oviposited on pyrethrum leaves and flower peduncles, but found that only a small fraction of those eggs hatched. Larvae were severely stunted in development, most likely due to the presence of pyrethrin defense compounds. This example of exploitative mutualism, which blocks the reproductive success of the moth pollinator and depends on nocturnal interactions, is placed into an ecological context to explain why it may have developed.

Transcriptomic Analysis of the CNL Gene Family in the Resistant Rice Cultivar IR28 in Response to Ustilaginoidea virens Infection.

Rice false smut, caused by Ustilaginoidea virens, threatens rice production by reducing yields and contaminating grains with harmful ustiloxins. However, studies on resistance genes are scarce. In this study, the resistance level of IR28 (resistant cultivar) to U. virens was validated through artificial inoculation. Notably, a reactivation of resistance genes after transient down-regulation during the first 3 to 5 dpi was observed in IR28 compared to WX98 (susceptible cultivar). Cluster results of a principal component analysis and hierarchical cluster analysis of differentially expressed genes (DEGs) in the transcriptome exhibited longer expression patterns in the early infection phase of IR28, consistent with its sustained resistance response. Results of GO and KEGG enrichment analyses highlighted the suppression of immune pathways when the hyphae first invade stamen filaments at 5 dpi, but sustained up-regulated DEGs were linked to the 'Plant-pathogen interaction' (osa04626) pathway, notably disease-resistant protein RPM1 (K13457, CNLs, coil-coiled NLR). An analysis of CNLs identified 245 proteins containing Rx-CC and NB-ARC domains in the Oryza sativa Indica genome. Partial candidate CNLs were shown to exhibit up-regulation at both 1 and 5 dpi in IR28. This study provides insights into CNLs' responses to U. virens in IR28, potentially informing resistance mechanisms and genetic breeding targets.

First Report of Anthracnose Caused by Colletotrichum jinshuiense on Goldthread in China.

Goldthread (Coptis chinensis Franch) is one of the most widely used Chinese traditional medicine plants with remarkable medicinal properties (Mizrahi et al. 2014). In July 2019, a new anthracnose-like leaf spot disease was observed in Banqiao Town, Enshi, Hubei, China. The incidence rate ranged from 10% to 20%. Infected leaves firstly showed oil-like dots, further gradually expanded to irregular whorls with a pale center and dark-brown edge. Petiole infection led to leaves dropping when severe occurrence. Black acervuli were developed on the infected leaves with abundant setae, especially near veins. To identify the causal agent, 4-mm2 tissues were derived from the disease-health junction and surface-disinfected with 0.1% mercury dichloride for 1 min and 75% ethanol for 30 s respectively. They were placed on a PDA plate and incubated at 25°C after being rinsed with sterile water three times. Isolates were purified by single spore isolation. Colonies on PDA were white to pale-gray with dense aerial mycelia, and the underside was yellowish to olive. Colonies grow 77.5 to 81.5 mm in 1 week. No conidia were observed during vegetable growth, but conidiomatal acervuli were found on infected leaves. Setae were 1-3 septate, dark-brown, 78.0 to 134.5 µm (mean = 108 ± 23.4) long, 4.1 to 9.1 µm (mean = 6.1 ± 1.1) diameter, cylindrical to conical, apices acute. Conidiophores hyaline to pale brown, septate. Conidia were hyaline, unicellular, aseptate, curved, cylindrical, often guttulate, measuring 20.1 to 28.0 × 3.5 to 5.4 µm (mean = 25.4 ± 1.7 × 4.5 ± 0.5 µm), L/W ratio = 5.6. Hyphae septate branched, hyaline to pale brown, 1.6 to 4.5 in diameter. Hyphopodial appressoria pale to medium brown, smooth-walled, globose or obovoid, 6.3 to 9.9 × 4.1 to 7.6 µm (mean = 8.3 ± 0.9 × 7.6 ± 0.7 µm), L/W ratio = 1.1. Morphological features were similar to the description of C. jinshuiense (Fu et al. 2019). To identify its phylogenetic position, maximum-likelihood (ML) analyses of two isolates (Esh8 and Esh 11) were implemented with a concatenation of multiple sequences of the internal transcribed spacer region (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), beta-tubulin (TUB2), and chitin-synthase 1 (CHS-1) using MEGA 7. The sequences were amplified using primers ITS1/ITS4, GDF1/GDR1, ACT-512F/ACT-783R, T1/Bt2b, CHS-79F/CHS-354R (Weir et al. 2012) and deposited in GenBank with accession numbers MW440484 - MW440485 (ITS), MW676256 - MW676257 (GAPDH), MW676252 - MW676253 (ACT), MW676254 - MW676255 (TUB2) and MW676258 - MW676259 (CHS-1). Results indicated they were clustered with C. jinshuiense in the C. dematium species complex. Isolates were inoculated onto injured healthy leaves (20 leaves) with mycelial plugs, ten leaves being inoculated with blank plugs were used as control. Disease symptoms were consistent with those observed in the field after five days post-inoculation with a 100% incidence rate, while no symptom was observed on the control leaves. And same isolates were isolated from six inoculated leaves with 100% re-isolation frequency. These results fulfilled Koch's postulates. In a previous study, C. boninense was identified as the causal agent of goldthread anthracnose in Chongqing, China (Ding et al. 2020). To our knowledge, this study is the first report of anthracnose on goldthread caused by C. jinshuiense in China.

Insight Into the Ultrastructure of Antennal Sensilla of Mythimna separata (Lepidoptera: Noctuidae).

The oriental armyworm, Mythimna separata (Walk), is one of the most serious pests of cereals in Asia and Australasia. The structure and distribution of the antennal sensilla of M. separata were studied by scanning electron microscopy and transmission electron microscopy. The results showed that antennae of both female and male M. separata are filiform in shape. Three groups and seven morphological sensillum types were recorded in both sexes, including uniporous sensilla (sensilla chaetica), multiporous sensilla (sensilla trichodea, basiconica, coeloconica, and styloconica), and aporous sensilla (sensilla squamiformia and Böhm bristles). S. trichodea, which were the most abundant sensilla, was made of three subtypes (ST I, ST II, and ST III) according to external features and two subtypes of s. basiconica (SB I and SB II) and s. coeloconica (SCo I and SCo II) were identified, respectively. Sexual dimorphisms in sensilla of M. separata were mainly perceived as the variations in the numbers of several sensilla subtypes. Also, the possible functions of the antennal sensilla were discussed. These results contribute to our understanding of the function of antennae in the behavior of M. separata.

Transcriptomic Analysis of Resistant and Wild-Type Isolates Revealed Fludioxonil as a Candidate for Controlling the Emerging Isoprothiolane Resistant Populations of Magnaporthe oryzae.

The point mutation R343W in MoIRR, a putative Zn2Cys6 transcription factor, introduces isoprothiolane (IPT) resistance in Magnaporthe oryzae. However, the function of MoIRR has not been characterized. In this study, the function of MoIRR was investigated by subcellular localization observation, transcriptional autoactivation test, and transcriptomic analysis. As expected, GFP-tagged MoIRR was translocated in the nucleus, and its C-terminal could autonomously activate the expression of reporter genes HIS3 and α-galactosidase in absence of any prey proteins in Y2HGold, suggesting that MoIRR was a typical transcription factor. Transcriptomic analysis was then performed for resistant mutant 1a_mut (R343W), knockout transformant ΔMoIRR-1, and their parental wild-type isolate H08-1a. Upregulated genes in both 1a_mut and ΔMoIRR-1 were involved in fungicide resistance-related KEGG pathways, including the glycerophospholipid metabolism and Hog1 MAPK pathways. All MoIRR deficiency-related IPT-resistant strains exhibited increased susceptibility to fludioxonil (FLU) that was due to the upregulation of Hog1 MAPK pathway genes. The results indicated a correlation between FLU susceptibility and MoIRR deficiency-related IPT resistance in M. oryzae. Thus, using a mixture of IPT and FLU could be a strategy to manage the IPT-resistant populations of M. oryzae in rice fields.

Climate warming promotes pesticide resistance through expanding overwintering range of a global pest.

Climate change has the potential to change the distribution of pests globally and their resistance to pesticides, thereby threatening global food security in the 21st century. However, predicting where these changes occur and how they will influence current pest control efforts is a challenge. Using experimentally parameterised and field-tested models, we show that climate change over the past 50 years increased the overwintering range of a global agricultural insect pest, the diamondback moth (Plutella xylostella), by ~2.4 million km2 worldwide. Our analysis of global data sets revealed that pesticide resistance levels are linked to the species' overwintering range: mean pesticide resistance was 158 times higher in overwintering sites compared to sites with only seasonal occurrence. By facilitating local persistence all year round, climate change can promote and expand pesticide resistance of this destructive species globally. These ecological and evolutionary changes would severely impede effectiveness of current pest control efforts and potentially cause large economic losses.

A chemosensory protein MsepCSP5 involved in chemoreception of oriental armyworm Mythimna separata.

Chemosensory proteins (CSPs) have been suggested to perform several functions in insects, including chemoreception. To find out whether MsepCSP5 identified from Mythimna separata shows potential physiological functions in olfaction, gene expression profiles, ligand-binding experiments, molecular docking, RNA interference, and behavioral test were performed. Results showed that MsepCSP5 was highly expressed in female antennae. MsepCSP5 showed high binding affinities to a wide range of host-related semiochemicals, and displayed that 26 out of 35 candidate volatiles were highly bound (Ki < 10 µM) at pH 5.0 rather than pH 7.4. The binding sites of MsepCSP5 to candidate volatiles were well predicted by three-dimensional structure modeling and molecular docking experiments. Pursuing further, biological activities of M. separata to highly bound compounds elicited strong behavioral responses, such as alcoholic compounds displayed strong attractiveness whereas terpenes showed repellency to M. separata. The transcript expression level of MsepCSP5 gene significantly decreased after injecting target dsRNAs, and resulted in non-significant preference responses of M. separata to semiochemicals, such as 3-pentanol and 1-octene-3-ol. In conclusion, MsepCSP5 may involve in semiochemical reception of M. separata.

De novo analysis of the oriental armyworm Mythimna separata antennal transcriptome and expression patterns of odorant-binding proteins.

To better understand the olfactory mechanisms in the oriental armyworm Mythimna separate, one of the most serious pests of cereals, an antennal transcriptome was constructed in this study. A total of 130 olfactory related transcripts were identified. These transcripts were predicted to encode 32 odorant-binding proteins (OBPs), 16 chemosensory proteins (CSPs), 71 olfactory receptors (ORs), 8 ionotropic receptors (IRs), 1 gustatory receptor (GR) and 2 sensory neuron membrane proteins (SNMPs). Q-PCR analysis of the temporal expression profiles of seven OBPs in different tissues indicated that, except for MsepOBP19 which was highly expressed in the wings of 0-day-old adult and MsepOBP20 which was low expressed in all tissues, other tested MsepOBPs were significantly more highly expressed in the antenna than in the head (antenna excluded), thorax, abdomen, legs and wings. The expression levels of MsepOBPs were diverse in different life stages (differed on eclosion days). MsepOBP5 exhibited female-biased expression in 0- and 5-day-old adult, while no gender bias in 1- and 3-day-old adult was detected and similar expression profiles were found for MsepOBP7, 20, 24 and 26. In addition, we found that although the expression of MsepOBP22 was female biased in 0- and 5-day-old adult, in the 3-day-old adult it was male-biased. Our findings established a foundation for future studies of the functions of olfactory proteins in M. separata.

Impact of hot events at different developmental stages of a moth: the closer to adult stage, the less reproductive output.

Hot days in summer (involving a few hours at particularly high temperatures) are expected to become more common under climate change. How such events at different life stages affect survival and reproduction remains unclear in most organisms. Here, we investigated how an exposure to 40 °C at different life stages in the global insect pest, Plutella xylostella, affects immediate survival, subsequent survival and reproductive output. First-instar larvae showed the lowest survival under heat stress, whereas 3rd-instar larvae were relatively heat resistant. Heat exposure at the 1(st)-instar or egg stage did not influence subsequent maturation success, while exposure at the 3rd-instar larval stage did have an effect. We found that heat stress at developmental stages closer to adult stage caused greater detrimental effects on reproduction than heat stress experienced at earlier life stages. The effects of hot events on insect populations can therefore depend critically on the timing of the event relative to an organism's life-cycle.

[Thermal tolerance of diamondback moth Plutella xylostella].

Diamondback moth Plutella xylostella is a worldwide important pest on cruciferous vegetables. Critical thermal maximum (CTMax) is often used as an index for the thermal tolerance of insects. By the method of dynamic heating, this paper measured the CTMax of P. xylostella in a self-assembled device, and studied the effects of development stage, rearing temperature, generation, sex, and heat shock on the thermal tolerance of P. xylostella based on the CTMax values. Reared at 25 degrees C, the mean CTMax of the 4th larva (50.31 degrees C) was significantly higher than that of the 1st larva (43.03 degrees C), 2nd larva (46.39 degrees C), 3rd larva (49.67 degrees C), female adult (45.76 degrees C), and male adult (47.73 degrees C); reared at 20, 25, and 30 degrees C, the adults had no significant difference in their CTMax; reared at 30 degrees C for 1-, 3-, and 6 generations, the CTMax of the adults also had no significant difference. In all the treatments, the CTMax of the female and male adults had less difference. Heat shock with 40 degrees C for 45 minutes could make the CTMax of 5 day-old male moth increased from 45.51 degrees C to 46.49 degrees C.

[Niches of dominant grub populations in different athletic areas of golf course].

The study on the temporal niche, spatial niche and spatial distribution of dominant grub populations in different athletic areas of golf courses in subtropics showed that except the lower temporal niche breadths of individual grubs in greens III and IV, the dominant grub populations in 4 areas of fairways and greens all had a >0.9 temporal niche breadth. The temporal niche breadth of 4 grub species in fairway highland and greens I and II was the highest, and that in rough highland and green IV was the lowest. For the grub species except Holotrichi-avata, the one-sided low vertical spatial niche breadth was 0.2-0.7. The temporal niche overlap in fairways and greens of 4 grub species was above 0.8, and the spatial niche overlap of great majority grub species was about 0.1-0.6. The spatial niche overlap between Anomala corpulenta and Anomala cupripes in fairways was the highest (>0.8), and that between Anomala corpulenta and Anomala cupripes and between Holotrichi asauteri and Holotrichi-avata in greens was the highest (>0.8). 4 grub species in fairways and greens mainly activated in 0-5 and 5-10 cm soil layers, a few grubs in 10-15 cm soil layer of fairway, and 4 grub species almost could not activate and feed in 10-15 cm soil layer of green.