Recognition of floral homeotic MADS domain transcription factors by a phytoplasmal effector,phyllogen, induces phyllody

Plant pathogens alter the course of plant developmental processes, resulting in abnormal morphology in infected host plants. Phytoplasmas are unique plant‐pathogenic bacteria that transform plant floral organs into leaf‐like structures and cause the emergence of secondary flowers. These distinctive symptoms have attracted considerable interest for many years. Here, we revealed the molecular mechanisms of the floral symptoms by focusing on a phytoplasma‐secreted protein, PHYL1, which induces morphological changes in flowers that are similar to those seen in phytoplasma‐infected plants. PHYL1 is a homolog of the phytoplasmal effector SAP54 that also alters floral development. Using yeast two‐hybrid and in planta transient co‐expression assays, we found that PHYL1 interacts with and degrades the floral homeotic MADS domain proteins SEPALLATA3 (SEP3), APETALA1 (AP1) and CAULIFLOWER (CAL). This degradation of MADS domain proteins was dependent on the ubiquitin–proteasome pathway. The expression of floral development genes downstream of SEP3 and AP1 was disrupted in 35S::PHYL1 transgenic plants. PHYL1 was genetically and functionally conserved among other phytoplasma strains and species. We designate PHYL1, SAP54 and their homologs as members of the phyllody‐inducing gene family of ‘phyllogens’.     

Efficacy of cashew kernel extracts in the control of the maize weevil,Sitophilus zeamais (Coleoptera: Curculionidae)

Six different solvents namely: methanol, ethanol, water, acetone, pet-ether or n-hexane was used to extract the essential oils from cashew kernel. The extracted oils were evaluated on Sitophilus zeamais (Motschulsky 1855) for mortality, oviposition and adult emergence effects. The long-term storage and water absorption capacity of the treated maize grains was also investigated. Results showed that the steam distillate was most lethal on S. zeamais. The least potent extracts on S. zeamais were those extracted with methanol and ethanol. The extracts drastically reduced oviposition in S. zeamais. Maize grains treated with steam distillate extract had the lowest water absorption capacity, while the highest water absorption capacity was obtained in grains treated with ethanol and methanol. Extracts of steam distillate, n-hexane, petroleum ether and acetone completely prevented infestation and subsequence damage of the treated maize grains for a period of three months. In the grains treated with the methanol and ethanol extracts, damage of 28.61% and 6.24%, respectively were obtained. The results obtained reveal that extracts of cashew kernel are effective in controlling S. zeamais and could serve as an alternative to synthetic insecticides.     

Elimination ofSitotroga cerealella (Olivier) bySitophilus zeamais Motschulsky from mixed laboratory cultures on maize

This paper describes a series of experiments conducted to determine why Sitophilus zeamaisMots . and Sitotroga cerealella (Oliv. ) could not survive together in maize cultures in the laboratory. The effect of S. zeamais on different developmental stages of S. cerealella was investigated. The presence of adult S. zeamais slightly affected moth copulation, egg laying and moth eggs in a mixed culture, but large numbers of developing moths inside maize grains were killed by the adult weevil through feeding on the grains. The major cause of elimination of S. cerealella by S. zeamais from mixed cultures was therefore found to be damage to the immature moths in grain and such moth mortality increased as the developing moths became bigger in the grains. A weevil: grain ratio of approximately 1.4∶1 was found to be the critical weevil density at which the moth disappeared from the mixed cultures.     

Constitutive expression of the SAP1 gene from willow (Salix discolor) causes early flowering in Arabidopsis thaliana

SAP1-1 and SAP1-2 were isolated from the male reproductive buds of willow (Salix discolor, clone S365). SAP1-1 differs from SAP1-2 based on a few nucleotide substitutions, but the sizes of their full-length cDNAs are identical. The deduced amino acid sequences of SAP1-1 and SAP1-2 were 98% similar and contain the same C-terminal amino acid motif “GYGA” like that of PTAP1-2 from Populus trichocarpa. The expression patterns of SAP1 in various parts of the male reproductive buds of S. discolor implicate this gene in the formation of the inflorescence meristems, bracts, and floral meristems. To characterize the functions of SAP1, we assessed Arabidopsis thaliana transformed with 35S∷SAP1-1. A total of 52 transgenic T₁ lines were obtained, and a 3:1 segregation ratio was obtained in the T₂ generation of each line. In the T₃ generation, five homozygous transgenic lines were obtained, which were used for further analysis. Screening of transgenic lines was greatly facilitated by the detection of GFP expression starting with germinating seeds. Phenotypes of the homozygous transgenic lines included early flowering, conversion of inflorescence branches to solitary flowers, formation of terminal flowers, and formation of flowers with greater number of petals, stamens, and pistils. Northern analysis showed similar expression levels in all five lines. This study provides the first functional analysis of an APETALA1 (AP1)/SQUAMOSA (SQUA) homolog from a dioecious species and suggests that SAP1 is a homolog of the AP1/SQUA gene.     

Dynamics of infestation and losses of stored maize due to the larger grain borer (Prostephanus truncatus Horn) and maize weevils (Sitophilus zeamais Motschulsky)

The dynamics of infestation and losses of stored maize due to the larger grain borer (Prostephanus truncatus Horn) and maize weevils (Sitophilus zeamais Motschulsky) were investigated in Tanzania. In single species populations, significantly higher maize damage (P ≤ 0.05) was caused by P. truncatus (23.3%) compared to S. zeamais (6.4%). Weight losses caused by P. truncatus were higher (12.7%) than for S. zeamais (6.7%) under similar conditions. A higher proportion of P. truncatus in the initial infestation population led to significantly higher (≤0.05) maize damage compared to equal or higher proportions of S. zeamais. Irrespective of the initial number of S. zeamais, population size increased to levels higher than for P. truncatus. Maize losses in unimproved storage structures were >80% compared to <2.5% in improved structures under natural infestation. Inter-specific competition appeared to influence the dynamics of infestation, damage levels and weight losses. The first two months of storage were critical for damaging insect populations to build up.     

Interspecific competition between Sitophilus zeamais and Sitotroga cerealella in a patchy environment

The maize weevil, Sitophilus zeamais (Motchulsky) (Coleoptera: Curculionidae), and the Angoumois grain moth, Sitotroga cerealella (Olivier) (Lepidoptera: Gelechiidae), are important insect pests of stored products. The coexistence in nature of the two species on maize has been difficult to explain from laboratory set‐ups, as the moth is inevitably eliminated by S. zeamais. However, early laboratory experiments ignored the spatial dimension, while several studies have lately revealed that two competing species may coexist in a spatially divided environment even though one of the species is competitively superior in a simpler universe. Two hypotheses for coexistence were examined by studying the interactions in a system where the resources were spatially divided. It was found that spatial subdivision in the current experiment was insufficient to ensure coexistence between the two species, so neither of the hypotheses could be confirmed. A simple model describing the population dynamics of S. zeamais was fitted to the data. Its parameters provide important information about how resource depletion influences the per capita growth rate. Finally, the study indicates that the mechanism responsible for S. zeamais's rapid elimination of S. cerealella is the superiority of the former species to colonize and monopolize new patches. The design of this study may serve as a template for further laboratory experiments revealing the effect of a spatially divided environment on competitive interactions. However, the present study also indicates that choosing the appropriate spatial scale may be critical for relating the results to natural environments.     

Development of a PCR-based marker to identify rice blast resistance gene,Pi-2(t), in a segregating population

The genomic clone RG64, which is tightly linked to the blast resistance gene Pi-2(t) in rice, provides means to perform marker-aided selection in a rice breeding program. The objective of this study was to investigate the possibility of generating a polymerase chain reaction (PCR)-based polymorphic marker that can distinguish the blast resistant gene, Pi-2(t), and susceptible genotypes within cultivated rice. RG64 was sequenced, and the sequence data was used to design pairs of specific primers for (PCR) amplification of genomic DNA from rice varieties differing in their blast disease responsiveness. The amplified products, known as sequenced-tagged-sites (STSs), were not polymorphic between the three varieties examined. However, cleavage of the amplified products with the restriction enzyme HaeIII generated a polymorphic fragment, known as specific amplicon polymorphism (SAP), between the resistant and the susceptible genotypes. To examine the power of the identified SAP marker in predicting the genotype of the Pi-2 (t) locus, we determined the genotypes of the F₂ individuals at this locus by performing progeny testing for the disease response in the F₃ generation. The results indicated an accuracy of more than 95% in identifying the resistant plants, which was similar to that using RG64 as the hybridization probe. The identification of the resistant homozygous plants increased to 100% when the markers flanking the genes were considered simultaneously. These results demonstrate the utility of SAP markers as simple and yet reliable landmarks for use in marker-assisted selection and breeding within cultivated rice.     

Efficacy of intercropping and harvest time modification for reducing field infestation of maize by Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae)

Most management practices of Sitophilus zeamais Motschulsky, a field-to-post-harvest insect pest of cereals, have focused on post harvest control methods. This experiment was designed to investigate the potential of cropping system and modification of time of harvest to control S. zeamais. Intercropping and harvest time modification had significant (P < 0.05) effect on the number of S. zeamais emerging 42 days post-harvest. For the early harvest (15 weeks after planting (WAP)), the mean number of S. zeamais recorded from a maize monoculture (7.39) was significantly (P < 0.05) higher than the mean numbers of weevils emerging from a maize–soybean intercrop (2.31), but not significantly higher than the number recorded in maize–groundnut (3.87) intercrop. For the late harvest (18 WAP), the mean number of emerged adult observed in the maize–soybean intercrop (6.13) was significantly lower than the mean number of adult emerging from the monocrop maize (13.24). Maize–groundnut intercrop did not significantly reduce field infestation of S. zeamais compared with monocrop maize. Percentage weight loss observed in early harvested maize was significantly (P < 0.0001) lower than what was observed in late-harvested maize. Percentage weight loss was highest in stored maize harvested from monocrop maize plots for the early harvest, whereas intercropping maize with soybean reduced percentage weight loss when harvest was delayed.     

Bioactivity of fenchone and Plectranthus glandulosus oil against Prostephanus truncatus and two strains of Sitophilus zeamais

The toxic, repellent and reproduction inhibitory effects of Plectranthus glandulosus essential oil and one of its major compounds, fenchone, were evaluated against adult Prostephanus truncatus (Horn) and for two strains of Sitophilus zeamais Motschulsky from Cameroon and Germany. Maize grains were coated with the oil at 0, 10, 20, 40 and 80 μl/40 g grain and fenchone at 0, 20, 40, 80 and 120 μl/40 g grain. Mortality was assessed over a 7‐day period, followed by the determination of F₁ progeny production. The repellence of the two products was evaluated at five rates (0.5, 1, 3, 5, 10 μl/half disc) in a choice bioassay on filter paper. The oil and fenchone caused significant dose‐dependent mortality to the three insects, with the oil achieving 100% and 90% mortality for the two S. zeamais strains and P. truncatus, respectively, within 1 day of exposure at the dosage of 80 μl/40 g grain. The insects were less susceptible to fenchone compared with the crude oil. The German strain of S. zeamais was susceptible to the oil than the Cameroonian strain. At the dosage of 20 μl/40 g grain, S. zeamais F₁ progeny emergency was completely inhibited by the oil, while 80 μl/40 g grain treatment with fenchone prevented also insect emergence. Plectranthus glandulosus oil generally evoked a maximum per cent repellency of 100% for the three insects, but fenchone caused 55–60% to S. zeamais and 80% to P. truncatus. These results suggest that fenchone, though a major constituent of P. glandulosus oil, may only be a minor component of its bioactivity.     

Contact and fumigant toxicities of calamusenone isolated from Acorus gramineus rhizome against adults of Sitophilus zeamais and Rhizopertha dominica

Abstract Calamusenone [3,8‐dimethyl‐5‐(1‐methylethylidene)‐1,2,3,4,5,6,7,8‐octahydroazulene‐6‐one, C₁₅H₂₂O] from Acorus gramineus Soland rhizome was tested in the laboratory for its insecticidal activities against adults of Sitophilus zeamais Motsch. and Rhizopertha dominica (Fab.), using dry film contact and fumigation methods. Responses varied with insect species, dosage and exposure time. In the dry film contact experiment, the highest insecticidal effects of calamusenone against S. zeamais and R. dominica adults were produced at 170.32 μg/cm² after treatment for 72 h, with 96.2% and 98.7% mortalities, respectively. The median lethal concentration (LC₅₀) (72 h) values of calamusenone against S. zeamais and R. dominica adults were 67.00 μg/cm² and 77.30 μg/cm², respectively. As a potential fumigant, calamusenone showed moderate insecticidal effect on the adults of S. zeamais and R. dominica in fumigation experiment, with their LC₅₀ (120 h) values of 125.71 μL/L and 93.64 μL/L respectively. Calamusenone isolated from A. gramineus rhizome showed promise as a novel pesticide candidate for stored‐product pest control.     

Suppression of the stored-product pest Sitophilus zeamais (Coleoptera: Curculionidae) by simultaneous release of a predatory bug and a parasitoid wasp

The control of the maize weevil, Sitophilus zeamais in brown rice was estimated by using a reduviid bug, Amphibolus venator, and a pteromalid wasp, Theocolax elegans. Results showed that the simultaneous release of both species of natural enemies could enhance biological control compared with the release of either species alone.     

Locomotor activity of Sitophilus zeamais and Sitotroga cerealella on maize

Some aspects of the locomotor activity of two pests of stored maize, Sitophilus zeamais Mots. and Sitotroga cerealella (Oliv.) were studied in the laboratory using a light dependent resistor actograph. Activity of S. zeamais adults increased weekly for 4 weeks and virgin males were more active than virgin females of the same age. When the sexes were mixed activity was intermediate between that of the separate sexes for about 3 weeks. Observations made on adults of S. cereablella for 4 days showed a daily increase in moth activity. When S. zeamais and S. cerealella were observed together, activity increased initially to about twice the mean activity of the species separately. Over 24 hr observation periods, no overt rhythms of activity were recorded for either species but S. zeamais was slightly more active between 11 and 19 hr GMT while in S. cerealella periods of low and high activity appeared to alternate nearly every 4–7 hours.

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Résumé Quelques aspects de l'activité locomotrice de deux ennemis du mais stocke Sitophilus zeamais Mots. et Sitotroga cerealella Oliv. ont été étudiés au laboratoire avec un actographe à résistor fonctionnant à la lumière.L'activité des adultes de S. zeamais augmente chaque semaine pendant 4 semaines, et les mâles vierges sont plus actifs que les femelles vierges du même âge. Quand les sexes sont mélangés l'activité est intermédiaire à celle des sexes séparés pour 3 semaines.Les observations sur 4 jours des adultes de S. cerealella montre une augmentation quotidienne de l'activité.Quand S. zeamais et S. cerealella sont observés ensemble, l'activité commence par doubler par rapport à celle des espèces séparées. Avec des observations par périodes de 24 heures, aucun rythme clar d'activité n'a été enregistré pour ces espèces, mais S. zeamais est légèrement plus actif entre 11 h et 19 h GMT, tandis que les périodes de faible et forte activités de S. cerealella apparaissent alternativement toutes les 4 à 7 heures.

     

OsBP-73, a rice gene,encodes a novel DNA-binding protein with a SAP-like domain and its genetic interference by double-stranded RNA inhibits rice growth

The SAP domain is a recently defined DNA binding domain that forms a helix-extended-helix structure. SAP proteins have been implicated in nuclear architecture and/or RNA metabolism. In this paper, we describe the cloning and characterization of a rice gene, OsBP-73, encoding a 375 amino acid protein with a SAP-like domain. We identified the binding sequence of OsBP-73 by gel retardation assays and southwestern blotting. Northern blot analysis demonstrated that OsBP-73 is weakly expressed in root, leaf and immature seed. OsBP-73 gene expression was also examined by histochemical studies of transgenic rice plants carrying an OsBP-73 5/GUS reporter gene. The reporter gene is mainly expressed in the tissues with high cell division activities, such as root tip, stem node, panicle and immature seed. Genetic interference of OsBP-73 gene expression by double-stranded RNA strikingly inhibits the whole plant growth but does not affect the passage from the juvenile to adult phase. These results suggest that OsBP-73 may play an important role in the regulation of cell proliferation.     

Cleome monophylla essential oil and its constituents as tick (Rhipicephalus appendiculatus) and maize weevil (Sitophilus zeamais) repellents

The repellency of the essential oil of the shrubCleome monophylla (Family: Capparidaceae) and identified constituents of the oil were evaluated against the livestock tick,Rhipicephalus appendiculatus and the maize weevil,Sitophillus zeamais. In a tick climbing repellency bioassay, the oil ofC. monophylla exhibited repellency which, at the highest dose, was comparable to that of the commercial arthropod repellent N,N-diethyl toluamide (DEET). In a Y-tube olfactometer bioassay,C. monophylla oil showed higher or comparable repellency againstS. zeamais relative to DEET at all the doses tested. 14 Compounds were identified in theC. monophylla oil by GC, GC-MS and coinjection with authentic samples. Terpenolene was found to occur in largest quantity (14%) followed by 1-α-terpeneol (10%), pentacosane (9%), (α+β)-humulene (8%), phytol (5%) and 2-dodecanone (4%). The most repellent components againstR. appendiculatus andS. zeamais were 1-α-terpeneol and 2-dodecanone. The overall pattern of repellency activity of theC. monophylla constituents with respect to the two arthropods was, however, different. The potential ofC. monophylla in tick and maize weevil control is discussed.     

Chrm3 protects against acinar cell necrosis by stabilizing caspase-8 expression in severe acute pancreatitis mice model

Acinar cells in acute pancreatitis (AP) die through apoptosis and necrosis, the impacts of which are quite different. Early clinical interference strategies on preventing the progress of AP to severe acute pancreatitis (SAP) are the elimination of inflammation response and inhibition of necrosis. Muscarinic acetylcholine receptor M3 was encoded by Chrm3 gene. It is one of the best-characterized receptors of pancreatic β cells and regulates insulin secretion, but its function in AP remains unclear. In this study, we explored the function of Chrm3 gene in the regulation of cell death in l -arginine-induced SAP animal models. We found that Chrm3 was upregulated in pancreatitis, and we further confirmed the localization of Chrm3 resided in both pancreatic islets and acinar cell membranes. The reduction of Chrm3 decreased the pathological lesion of SAP and reduced amylase activities in serum. Consistently, Chrm3 can suppress acinar cells necrosis markedly, but has no effect on regulating apoptosis after l -arginine treatment. It was shown that Chrm3 attenuated acinar cells necrosis at least in part by stabilizing caspase-8. Thus, this study indicates that Chrm3 is critical participants in SAP, and regulation of Chrm3 expression might be a useful therapeutic strategy for preventing pathologic necrosis.     

Hexane extracts from Tephrosia vogelii Hook. f. protect stored maize against the weevil Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae)

Powdered dried leaves of Tephrosia vogelii (Hook) (Leguminosae: Papilionoideae) were extracted using hexane, acetone, and ethanol. The extracts were tested for their ability to protect stored maize from damage by Sitophilus zeamais Motschulsky, the major maize weevil in Africa. The acetone and ethanol extracts were ineffective, but the hexane extract had a relatively high efficacy, producing within 7 days a slow reduction in adult survival, reduced numbers of eggs laid and reduced numbers of F₁ progeny, resulting in seed damage averaging 8.8% compared with 98.6% in the untreated control after one generation. This hexane extract provided control of S. zeamais at a higher level compared with neem (the botanical control) but at a lower level compared with pirimiphos‐methyl (the synthetic control). These findings are discussed in the context of T. vogelii containing rotenoids.     

Characterization and identification of the candidate gene of rice thermo-sensitive genic male sterile gene <Emphasis Type="Italic">tms5</Emphasis> by mapping

Previous research has demonstrated that the thermo-sensitive genic male-sterile (TGMS) gene in rice was regulated by temperature. TGMS rice is important to hybrid rice production because the application of the TGMS system in two-line breeding is cost-effective, simple, efficient and overcomes the limitations of the cytoplasmic male sterility (CMS) system. AnnongS is the first discovered and deeply studied TGMS rice line in China. Previous studies have suggested that AnnongS-1 and Y58S, two derivative TGMS lines of AnnongS, were both controlled by a single recessive gene named tms5, which was genetically mapped on chromosome 2. In the current study, three populations (AnnongS-1 × Nanjing11, Y58S × Q611, and Y58S × Guanghui122) were developed to investigate the tms5 gene molecular map. Analysis of recombination events of sterile samples, utilizing 125 probes covering the tms5 region, suggested that the tms5 gene was physically mapped to a 19 kb DNA fragment between two markers, 4039-1 and 4039-2, located on the BAC clone AP004039. Following the construction of a physical map between the two markers, ONAC023, a member of the NAC (NAM-ATAF-CUC-related) gene family, was identified as the candidate of the tms5 gene.