In Situ Aquaculture Methods for Dysidea avara (Demospongiae,Porifera) in the Northwestern Mediterranean

Marine sponges produce secondary metabolites that can be used as a natural source for the design of new drugs and cosmetics. There is, however, a supply problem with these natural substances for research and eventual commercialisation of the products. In situ sponge aquaculture is nowadays one of the most reliable methods to supply pharmaceutical companies with sufficient quantities of the target compound. In this study, we focus on the aquaculture of the sponge Dysidea avara (Schmidt, 1862), which produces avarol, a sterol with interesting pharmaceutical attributes. The soft consistency of this species makes the traditional culture method based on holding explants on ropes unsuitable. We have tested alternative culture methods for D. avara and optimized the underwater structures to hold the sponges to be used in aquaculture. Explants of this sponge were mounted on horizontal ropes, inside small cages or glued to substrates. Culture efficiency was evaluated by determination of sponge survival, growth rates, and bioactivity (as an indication of production of the target metabolite). While the cage method was the best method for explant survival, the glue method was the best one for explant growth and the rope method for bioactivity.     

Glycosides from Marine Sponges (Porifera, Demospongiae): Structures, Taxonomical Distribution, Biological Activities and Biological Roles

Literature data about glycosides from sponges (Porifera, Demospongiae) are reviewed. Structural diversity, biological activities, taxonomic distribution and biological functions of these natural products are discussed.     

Indole Alkaloids of the Stigonematales (Cyanophyta): Chemical Diversity,Biosynthesis and Biological Activity

The cyanobacteria are well recognized as producers of a wide array of bioactive metabolites including toxins, and potential drug candidates. However, a limited number of taxa are generally considered with respect to both of these aspects. That said, the order Stigonematales, although largely overlooked in this regard, has become increasingly recognized as a source of bioactive metabolites relevant to both human and environmental health. In particular, the hapalindoles and related indole alkaloids (i.e., ambiguines, fischerindoles, welwitindolinones) from the order, represent a diverse, and phylogenetically characteristic, class of secondary metabolites with biological activity suggestive of potential as both environmental toxins, and promising drug discovery leads. The present review gives an overview of the chemical diversity of biologically active metabolites from the Stigonematales—and particularly the so-called hapalindole-type alkaloids—including their biosynthetic origins, and their pharmacologically and toxicologically relevant bioactivities. Taken together, the current evidence suggests that these alkaloids, and the associated cyanobacterial taxa from the order, warrant future consideration as both potentially harmful (i.e., “toxic”) algae, and as promising leads for drug discovery.     

Suppressing Mediterranean fruit fly (Diptera: Tephritidae) with an attract-and-kill device in pome and stone fruit orchards in Western Australia

Attract-and-kill is a possible alternative to the conventional control of Mediterranean fruit fly (medfly), Ceratitis capitata Wiedemann (Diptera: Tephritidae) with bait or cover sprays of organophosphate insecticides. In this study, we determined if an attract-and-kill device (MagMED^®) could be applied alone at the recommended rate (80 devices/ha), or needed to be combined with other conventional control tactics to suppress medfly in deciduous fruit orchards (apple, nectarine, peach, pear, plum). We also determined if attract-and-kill had any detrimental effects on naturally occurring beneficial insects. Based on an acceptable level of crop loss of 2–5% (the level of crop loss growers can tolerate), attract-and-kill suppressed medfly and reduced crop loss below the acceptable level in orchards with additional controls. Attract-and-kill was most effective when combined with either bait spraying, cover spraying or both. Combining attract-and-kill with cover sprays of fenthion resulted in a drop in crop loss of apples from 2.5% to 0.5%. By combining attract-and-kill with weekly bait sprays and two cover spray applications, crop loss fell from 13.7% to 4.3% in peaches, from 6.3% to 5% in nectarines and from 6.3% to 3.7% in pears. Twice weekly bait spraying was the most effective technique at suppressing medfly, providing acceptable control when applied alone, which further improved by combining with attract-and-kill. By combining attract-and-kill with twice weekly bait spraying crop loss fell from 2% to 1% in nectarines and from 4% to 2% in apples. In laboratory trials, attract-and-kill devices killed 100% of brown lacewings (Micromus tasmaniae Walker) and 94% of transverse ladybeetles (Coccinella transversalis Fabricius) 24 h after contact. However, orchard samples suggest that it may not pose any significant negative impact on naturally occurring beneficials such as Neuroptera, Hymenoptera, Coleoptera, Thysanoptera and Syrphidae in the field.     

Contribution of main stem and tillers to durum wheat (Triticum turgidum L. var. durum) grain yield and its components grown in Mediterranean environments

Under terminal drought conditions, cereal varieties with limited tillering have been suggested to be advantageous, because they have fewer nonproductive tillers, thereby limiting water consumption prior to anthesis. In this study, four field trials were conducted over two growing seasons in southern Spain, under rainfed and irrigated conditions. Twenty-five genotypes were studied to evaluate the contribution of the main stem (MS) and tillers to grain yield and its components. Significant differences were found among genotypes for these contributions under non-stressed environments, but these differences were not significant under water-stress conditions. The contribution of the MS to plant grain yield was higher than that of tillers (68% vs. 32%) and was stable between years in irrigated trials. However, in the rainfed trials, MS contributed differently depending on year-to-year climate variations. Thus, under favorable weather conditions the contribution of MS to grain yield was higher than in the unfavorable year (85% vs. 59%). In irrigated environments, MS and tiller grain yield depended on the number of grains per spike, spikelets per spike, and thousand kernel weight (TKW). Under water-limited conditions, MS yield depended on the number of grains per spike and grains per spikelet, whereas the number of spikelets and TKW had less influence on MS grain yield. Furthermore, under water-stress conditions, high tillering genotypes showed yield levels similar to the genotypes with restricted tillering. Additionally, there was no significant evidence of a positive or negative effect of maximum tiller number on grain yield under rainfed conditions.     

橡胶树多主棒孢拮抗细菌BACYitc8-7的分离鉴定及其挥发性抑菌物质组分分析

多主棒孢菌（Corynespora cassiicola）是为害橡胶叶片的一种重要病原真菌,目前生产上普遍使用化学农药防治该病害,给植胶区的生态环境带来很多负面影响,从橡胶树根际土壤中筛选对橡胶树多主棒孢菌具有高拮抗活性的细菌,可为橡胶树棒孢霉落叶病生防药剂的研发提供理论依据。本研究通过稀释涂布、平板对峙法从发病橡胶树根际土壤中分离筛选到1株对橡胶树多主棒孢菌具有高拮抗活性的细菌,抑菌带为(57.68±0.66)mm,且该菌株对9种重要的植物病原真菌均具有抑制作用;经过形态观察、生理生化检测及16S rRNA基因序列分析,结果发现其与铜绿假单孢菌基本相符,命名为BACYitc8-7;以LB、NA、高氏、金氏B和PDA为营养来源,评定不同营养条件下菌株BACYitc8-7挥发性物质对多主棒孢菌的抑制作用,发现菌株BACYitc8-7在金氏B和LB培养基上的拮抗作用最强,抑菌率达43.47%~48.69%;利用顶空固相微萃取-气相色谱-质谱法（HS-SPME-GC-MS）收集并检测菌株BACYitc8-7挥发性物质的抑菌组分,获得包括胺类、醇类、烯类、酚类、酯类、吡嗪类等35种物质,其中含量前5的物质为胺、硼烷二甲硫醚络合物、异戊醇、1-十三烯和2-甲基-3-甲硫基呋喃。     

Evaluation of structural and antibiosis resistance mechanisms during selection against Mediterranean corn borer (Sesamia nonagrioides Lef) in the maize synthetic EPS12

In maize, the Mediterranean corn borer (MCB), Sesamia nonagrioides Lef, is the insect pest that causes the most significant yield losses in north-western Spain. A S₁ recurrent selection program to improve resistance of EPS12 against MCB was evaluated previously. In the current study two experiments were conducted to determine if antibiosis and/or structural mechanisms were responsible for the improvement in resistance in this synthetic. Larval growth or survival of MCB was not significantly affected to consider the presence of antibiotic factors in the pith. Besides, pith puncture resistance, previously reported as a possible resistance trait in single inbred lines, did not show significant differences between cycles of selection. Length of the internode basal ring (LIBR) was the structural character most related to stem resistance improvement to MCB in the current evaluation. There was a high positive correlation between MCB tunnel length and LIBR (P ≤ 0.01, R² = 0.99). The larvae would enter the plant throughout the internode basal area and, this way, they could avoid the increased thickness or hardness of the remaining rind area. LIBR might be a useful measure in future resistance breeding programs against this borer.     

Assessment of lure and kill and mass-trapping methods against the olive fly,Bactrocera oleae (Rossi), in desert-like environments in the Eastern Mediterranean

Management of the olive fly using environmentally friendly methods includes strategies based on lure and kill and/or mass-trapping. Despite a wealth of studies related to the efficacy of different lure and kill and mass-trapping systems in several olive producing areas, there are few known regarding the performance of such systems in low input olive farms of the desert-like areas of the Middle East. The present study reports on the control of the olive fly using both lure and kill and mass-trapping devices in low-input farms in the region of Tubas, Palestine, between 2010 and 2012. The effect of environmental factors, such as elevation, on general patterns of olive fly infestation and the lure and kill system's performance was also studied. Our final goal included the development of a general strategy to produce organic olive oil using alternative olive fly control methods. Initially we used a commercially available lure and kill device, the Eco-Trap^®. The Eco-Trap performed relatively well during the three years of study, significantly reducing olive fly damage levels. Eco-Trap effectiveness was more evident at elevation above 400 m, where damage in the region of Tubas was more intense than at lower elevations. In a subsequent trial we tested a low-cost mass-trapping device. This device demonstrated similar levels of protection to the one seen with the Eco-Trap. Results showed that lure and kill and mass-trapping tactics can be effectively applied in the region of Tubas, and that accessible devices can be adopted to reduce control costs. The results also showed differential levels of damage throughout the region, which were related to elevation above sea level. The possibility of applying a regional management concept throughout the area by a centralized organization, such as a farmer's union based on mass-trapping systems seems feasible and is discussed.     

灰飞虱对氟虫腈抗性风险评估、遗传分析及杀虫剂敏感性研究

 在室内采用稻茎浸渍法进行了灰飞虱对氟虫腈抗性风险评估、抗性遗传分析和杀虫剂的敏感性研究。于2005年采自无锡麦田的灰飞虱种群在室内饲养43代期间用氟虫腈筛选了41代,结果此灰飞虱种群的抗性从8.4倍上升到2305倍。根据Tabashnik介绍的方法计算,现实遗传力(h2)分别为0.0388(1~31代)、0.2636(32~43代)和0.1113(1~43代), 表明灰飞虱对氟虫腈具有一定的抗性风险。采用稻茎浸渍法测定了2007-2008年江苏无锡和浙江长兴两地灰飞虱种群对15种杀虫剂的敏感性, 结果表明,苯基吡唑类杀虫剂氟虫腈、丁烯氟虫腈和乙虫腈的毒力最高(LC50=0.2~1.7 mg/L),其次为烯啶虫胺、噻虫嗪、毒死蜱、吡蚜酮(LC50=1.7~9.7 mg/L),其余杀虫剂的毒力较低;按照年度间敏感性变化,发现2008年无锡灰飞虱种群对氟虫腈和乙虫腈的敏感性比2007年降低了1.1倍。近年来氟虫腈已广泛用于防治这种害虫,但乙虫腈几乎没有使用。因此,对氟虫腈敏感性降低的大田灰飞虱种群似乎对乙虫腈存在交互抗性。通过抗(R)、感(S)亲本、正反交(F1、 F1′)、自交(F2)及回交(BC)后代对氟虫腈的剂量反应研究了灰飞虱对氟虫腈的抗性遗传特性,结果表明,其抗性为常染色体的不完全显性遗传\[D(F1)=0.20, D(F1′)=0.38\],抗性由2个或2个以上等位基因控制。还对灰飞虱的抗性治理进行了讨论。     

CAP Analysis of the Distribution of the Introduced Bemisia tabaci (Hemiptera: Aleyrodidae) Species Complex in Xinjiang,China and the Southerly Expansion of the Mediterranean Species

Bemisia tabaci (Gennadius) cryptic complex has invaded Xinjiang, China, since 1998. The distribution of Mediterranean (MED) and Middle East-Asia Minor 1 (MEAM1) B. tabaci substrains has been gradually identified due to the development of molecular technology. In this study, the distribution of MED and MEAM1 in Xinjiang was determined by cleaved amplified polymorphic sequence (CAPs). Results showed that MED dominated in northern Xinjiang (84%), whereas MEAM1 was dominant in southern Xinjiang (72%). Five pairs of simple sequence repeat (SSR) primers were used to analyze the genetic diversity of B. tabaci among 36 geographic populations. The genetic diversity of MED and MEAM1was low and varied little among populations in Xinjiang (0.09 ± 0.14 and 0.09 ± 0.13, respectively). Based on ∆K statistic, 13 populations of MEAM1 could be classified into two subgroups at K = 2, whereas the 23 populations of MED could be classified into four subgroups at K = 4. However, Mantel t-test demonstrated no correlation between geographical and genetic distances among B. tabaci complex (R = 0.42, P = 1.00). Neighbor-joining and principal coordinate analysis showed that geographical isolation and interspecific differences were the main causes of the genetic variation. Gene flow predicted that MEAM1 was most likely introduced from Urumqi to the southern Xinjiang. Meanwhile, a large proportion of MED in Kashi region came from Changji and Yining. To block ongoing dispersal, strict detection and flower quarantine regulations need to be enforced.     

Identification and Expression of Two Novel Cytochrome P450 Genes,CYP6CV1 and CYP9A38, in Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)

Cnaphalocrocis medinalis Güenée can cause severe losses in rice. Cytochrome P450s play crucial roles in the metabolism of allelochemicals in herbivorous insects. Two novel P450 cDNAs, CYP6CV1 and CYP9A38, were cloned from the midgut of C. medinalis. CYP6CV1 encodes a protein of 500 amino acid residues, while CYP9A38-predicted protein has 531 amino acid residues. Both cDNA-predicted proteins contain the conserved functional domains for all P450s. Phylogenetic analyses showed that CYP6CV1 is grouped in the cluster containing CYP6B members, while CYP9A38 is in the cluster including CYP9 members. However, both clusters are contained in the same higher lineage. Homologous analysis revealed that CYP6CV1 is most similar to CYP6B8, CYP6B7, CYP6B6, CYP6B2, and CYP6B4 with the highest amino acid identity of 41%. CYP9A38 is closest to CYP9A17, CYP9A21, CYP9A20, and CYP9A19 with the highest amino acid identity of 66%. Studies of temporal expression profiles revealed that CYP9A38 showed a steady increase in mRNA level during the five instar stages, but a low-expression level in pupae, and then presented at a high-expression level again in adults. Similar expression patterns were obtained with CYP6CV1. In the fifth instar larvae, CYP6CV1 was mainly expressed in midgut and fat bodies, whereas CYP9A38 was mainly expressed in midgut. Expression studies also revealed a 3.20-fold over-expression of CYP6CV1 and 3.54-fold over-expression of CYP9A38 after larval exposure to host rice resistance. Our results suggest that both CYP6CV1 and CYP9A38 may be involved in detoxification of rice phytochemicals.     

Assessment of avoidable losses and economic injury levels for the sorghum head bug, Calocoris angustatus Leth. (Hemiptera:Miridae) in India

Avoidable losses and economic injury levels (EILs) were computed for the sorghum head bug, Calocoris angustatus Leth., on three cultivars during three rainy seasons (1985–1987) at ICRISAT. Grain yield in plots protected with 2–5 insecticide sprays (carbaryl at 500 g a.i./ha) between the half-anthesis and the dough stage was significantly higher than in untreated plots, with cost-benefit ratios > 1. Plots that were not protected at half-anthesis and/or complete-anthesis, and at milk and dough stages, suffered a significant loss in yield. Bug damage spoiled the grain quality in terms of germination, 1000-grain mass, grain hardness and percentage floaters. Three to four sprays between complete-anthesis and the dough stage prevented a significant reduction in grain quality. Head bug density at half-anthesis, complete-anthesis, milk and dough stages was significantly and negatively associated with grain yield. Bug density at different stages explained 43–94% of the variance in yield. Direct effects of bug numbers at the milk stage and the indirect effects of head bug density at other stages through the milk stage contributed towards maximum reduction in yield. Maximum avoidable losses were recorded in cultivar ICSV 1 (88·6%), followed by CSH 1 (69·9%), and CSH 5 (53·9% in 1986 and 55·0% in 1987). Loss in grain yield/ha due to one insect/panicle at half-anthesis and that based on natural increase, were 548 and 232 kg in CSH 5 during 1986 and 1987, respectively, 251 kg in ICSV 1, and 77 kg in CSH 1. EILs based on a cost-benefit ratio of 1:1 ranged from 1·3–1·4 insects/panicle for CSH 1, 0·4 for ICSV 1, and 0·4–0·6 and 0·2–0·4 for CSH 5 during 1986 and 1987, respectively. Simple cost estimates for insecticide sprays and the resultant saving in grain yield are a useful tool for decision-making in pest management.     

Impact of Agronomic Strategies (Seed Tuber Pre-sprouting,Cultivar Choice) to Control Late Blight (<Emphasis Type="Italic">Phytophthora infestans</Emphasis>) on Tuber Growth and Yield in Organic Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Crops

In order to optimize potato crop management in organic farming systems, knowledge of crop growth processes in relation to limitations and reductions by nitrogen (N) and disease is necessary. This paper examines the effect of different prevention measures (seed tuber pre-sprouting, choice of cultivars: resistance to Phytophthora infestans; earliness of tuber initiation) against disease-related constraints on yields, depending on the N-mediated growing conditions of organic potato crops. Under conditions of a relatively high N supply, accelerating the early development and tuber initiation of potato crops by seed tuber pre-sprouting (yield increase of c. 18–23%) or the selective choice of cultivars with an earlier tuber initiation (yield increase of c. 0–21%) are the most effective strategies in combating late blight. They bring forward the crop development c. 7–10 days, escaping the negative impact of the fungus on tuber growth. Under conditions of a relatively low N supply, these strategies do not affect final tuber yield. The reason for the differences of the effect depending on N supply of the crops is, that the lower the N supply, the shorter the period of time over which tuber growth takes place, independent of whether P. infestans becomes severe or not. Tuber growth of organic crops low in N has mostly ceased by the time late blight becomes potentially important for limiting yield, with the consequence that preventative measures against P. infestans are meaningless. Surprisingly, the level of cultivar haulm resistance to late blight did not affect tuber yields in years with an early and strong late blight development. Probably, the positive effect of a longer resistance to the fungus (c. 1 week) was counterbalanced by a later tuber initiation (also c. 1 week). In years with a moderate late blight incidence, less susceptible cultivars were able to produce higher tuber yields on fields with a lower N availability (c. 17–20%), probably due to higher N use efficiency and a better match of N mineralization and N uptake. Under conditions of a relatively high N supply, the tested cultivars with a higher haulm resistance to late blight were not able to improve tuber yields.