Contrasting patterns of genetic differentiation in Macaronesian lineages of Ilex (Aquifoliaceae)

Islands offer an interesting framework in which to study the effect of geographical isolation on population genetic differentiation. For plant species with high dispersal abilities, however, oceanic barriers may not represent a factor promoting strong population structure. In this work, we analysed seven nuclear microsatellite loci in Ilex (Aquifoliaceae), a bird‐dispersed plant group, to infer patterns of genetic differentiation among Macaronesian taxa: I. canariensis, I. perado ssp. lopezlilloi, I. perado ssp. platyphylla (Canary Islands) and I. perado ssp. azorica (Azores). In agreement with current taxonomic classification, our results revealed a high genetic differentiation between Ilex lineages (I. canariensis and the I. perado complex), and also supported previous hypotheses that these are the result of independent dispersal events to the islands. In contrast, genetic differentiation between I. perado ssp. azorica and the two subspecies from the Canaries was high, suggesting that taxonomic revision may be necessary. Levels of genetic variation at microsatellite loci in ssp. azorica were, in addition, the lowest reported among Macaronesian bird‐dispersed taxa. Lastly, low genetic differentiation was observed between subspecies occurring on the same island (sspp. platyphylla and lopezlilloi). In summary, our results revealed contrasting patterns between Macaronesian Ilex lineages: I. canariensis displayed moderate population structure across islands, whereas the I. perado complex showed strong differentiation among populations sampled on different islands. Thus, the Macaronesian Ilex taxa show that long‐distance dispersal syndromes (ornithochory) do not always ensure genetic connectivity across large areas in island systems. Plant groups that successfully colonized the islands on multiple occasions may have found barriers to gene flow within certain lineages. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 258–268.     

A spectrophotometric technique for measuring particle ingestion by black fly larvae

A spectrophotometric technique was developed to provide insight into the feeding behavior of Simulium vittatum Zetterstedt (Diptera: Simuliidae) larvae. Larvae were exposed to water insoluble Neon Red particles (NRP) (DayGlo^®) in a controlled current. The insoluble particles were available for capture by the cephalic fans of the larvae and subsequent ingestion. The length of gut occupied by the particles after a given exposure time was determined by visual inspection and measured with the aid of a dissecting microscope. Larvae were then homogenized in acetone to solubilize the particles. After filtration, the quantity of pigmented particles in the alimentary tract of the larvae was determined using spectrophotometric analysis. The quantity of particles per unit length of the alimentary tract was calculated. Experiments were conducted to determine the ideal concentration of NRP for obtaining an accurate measure of ingestion without interfering with normal larval feeding behavior. Larval mortality following ingestion of insecticidal proteins produced by Bacillus thuringiensis ssp. israelensis was used as an indirect measure of feeding behavior for these experiments. A concentration of 15 p.p.m. of NRP in the larval medium was the highest concentration used that did not interfere with larval mortality following exposure to the insecticidal proteins. Additional experiments demonstrated that components of the experimental matrix did not interfere with NRP absorbance. The final experiment revealed that the consumption of NRP and insecticidal proteins by larvae was influenced by clay and cellulose in the larval medium.     

不结球白菜种质资源对TuMV的抗性鉴定与评价

分别利用苗期人工接种鉴定及ELISA检测方法,对127份不结球白菜种质资源进行芜菁花叶病毒(TuMV)的抗性鉴定。结果显示,苗期人工接种鉴定的病情指数(DI)分布在3.55~95.68之间,不同种质间表现出较大的抗性差异。不同抗性级别的次数分布图基本符合正态分布,略向感病区域偏离。基于病情指数的聚类分析结果与抗病性分级基本一致。通过苗期鉴定共筛选获得高抗TuMV的不结球白菜种质6份,抗病种质13份,其主要农艺性状表现出一定的多样性。其中叶面皱缩和具有刺毛的抗病材料所占比例较高,可能与TuMV抗性存在一定的相关性。ELISA检测结果显示,117份供试种质的P/N值分布在3.10~25.37之间,不同种质间表现出病毒含量的差异,但与DI值未呈现明显相关性,可作为抗病材料筛选的辅助指标。     

Chemical Composition of Ballota macedonica Vandas and Ballota nigra L. ssp. foetida (Vis.) Hayek Essential Oils – The Chemotaxonomic Approach

The essential oils isolated from fresh aerial parts of Ballota macedonica (two populations) and Ballota nigra ssp. foetida were analyzed by GC and GC/MS. Eighty five components were identified in total; 60 components in B. macedonica oil (population from the Former Yugoslav Republic of Macedonia), 34 components in B. macedonica oil (population from the Republic of Serbia), and 33 components in the oil of B. nigra ssp. foetida accounting for 93.9%, 98.4%, and 95.8% of the total oils, respectively. The most abundant components in B. macedonica oils were carotol (13.7 – 52.1%), germacrene D (8.6 – 24.6%), and (E)‐caryophyllene (6.5 – 16.5%), while B. nigra ssp. foetida oil was dominated by (E)‐phytol (56.9%), germacrene D (10.0%), and (E)‐caryophyllene (4.7%). Multivariate statistical analyses (agglomerative hierarchical cluster analysis and principal component analysis) were used to compare and discuss relationships among Ballota species examined so far based on their volatile profiles. The chemical compositions of B. macedonica essential oils are reported for the first time.     

Worldwide distribution and theoretical spreading of Trichoferus campestris (Coleoptera: Cerambycidae) depending on the main climatic elements

The velvet longhorned beetle, Trichoferus campestris (Coleoptera: Cerambycidae) is a serious wood‐boring pest that is a major threat to the phytosanitary condition of forests and orchards. Its worldwide expansion is a major concern for plant health. We have collected all bibliographical references, phytosanitary reports and authentic photographic evidence from entomological websites to determine the worldwide distribution of T. campestris. The theoretical directions of the spreading and actually occupied area of this arthropod pest were determined over the whole Holarctic range. Furthermore, the potential distribution area was calculated using cumulated temperature in the growing season averaged over 15 years both in the Palearctic and Nearctic regions. Holarctic expansion of the species, including the main parts of North America and Eurasia, is clearly indicated. Its populations occur in 29 countries to date, supported by documentation from 64 publications and 30 online forums. Its spread is continuously westward in the Palearctic; in the Nearctic, the spread was first southward from the Great Lakes region then eastward from Utah. The species has excellently adapted to circumstances of freight by ship and plane, as wood is often used, ensuring optimal conditions for the pest. In addition, the active spreading achieved by flying is an important factor contributing to its expansion. The primary criterion for controlling the species would be the introduction of a monitoring system in affected and exposed areas.     

Exome sequences and multi‐environment field trials elucidate the genetic basis of adaptation in barley

Broadening the genetic base of crops is crucial for developing varieties to respond to global agricultural challenges such as climate change. Here, we analysed a diverse panel of 371 domesticated lines of the model crop barley to explore the genetics of crop adaptation. We first collected exome sequence data and phenotypes of key life history traits from contrasting multi‐environment common garden trials. Then we applied refined statistical methods, including some based on exomic haplotype states, for genotype‐by‐environment (G×E) modelling. Sub‐populations defined from exomic profiles were coincident with barley's biology, geography and history, and explained a high proportion of trial phenotypic variance. Clear G×E interactions indicated adaptation profiles that varied for landraces and cultivars. Exploration of circadian clock‐related genes, associated with the environmentally adaptive days to heading trait (crucial for the crop's spread from the Fertile Crescent), illustrated complexities in G×E effect directions, and the importance of latitudinally based genic context in the expression of large‐effect alleles. Our analysis supports a gene‐level scientific understanding of crop adaption and leads to practical opportunities for crop improvement, allowing the prioritisation of genomic regions and particular sets of lines for breeding efforts seeking to cope with climate change and other stresses.     

A novel 3-oxoacyl-ACP reductase (FabG3) is involved in the xanthomonadin biosynthesis of Xanthomonas campestris pv. campestris

Xanthomonas campestris pv. campestris (Xcc), the causal agent of black rot in crucifers, produces a membrane-bound yellow pigment called xanthomonadin to protect against photobiological and peroxidative damage, and uses a quorum-sensing mechanism mediated by the diffusible signal factor (DSF) family signals to regulate virulence factors production. The Xcc gene XCC4003, annotated as Xcc fabG3, is located in the pig cluster, which may be responsible for xanthomonadin synthesis. We report that fabG3 expression restored the growth of the Escherichia coli fabG temperature-sensitive mutant CL104 under non-permissive conditions. In vitro assays demonstrated that FabG3 catalyses the reduction of 3-oxoacyl-acyl carrier protein (ACP) intermediates in fatty acid synthetic reactions, although FabG3 had a lower activity than FabG1. Moreover, the fabG3 deletion did not affect growth or fatty acid composition. These results indicate that Xcc fabG3 encodes a 3-oxoacyl-ACP reductase, but is not essential for growth or fatty acid synthesis. However, the Xcc fabG3 knock-out mutant abolished xanthomonadin production, which could be only restored by wild-type fabG3, but not by other 3-oxoacyl-ACP reductase-encoding genes, indicating that Xcc FabG3 is specifically involved in xanthomonadin biosynthesis. Additionally, our study also shows that the Xcc fabG3-disrupted mutant affects Xcc virulence in host plants.     

盐及干旱胁迫对油菜抗氧化系统和RbohC、RbohF基因表达的影响

以白菜型油菜‘陇油6号’和‘天油2号’为试验材料,经MAPK抑制剂U0126、H_2O_2清除剂DMTU、NADPH氧化酶抑制剂DPI和IMD预处理后再分别进行盐胁迫、PEG-6000模拟干旱胁迫,研究其对两种油菜幼苗活性氧、抗氧化酶活性和RbohC、RbohF基因表达的影响.结果表明:盐胁迫和PEG-6000模拟干旱胁迫下,两种白菜型油菜中H_2O_2积累量上升,O_2^-·积累量下降,抗氧化酶(超氧化物歧化酶SOD、过氧化氢酶CAT、抗坏血酸过氧化物酶APX和谷胱甘肽还原酶GR)活性和RbohC、RbohF基因表达均升高.与单独胁迫处理相比,两种油菜O_2^-·积累、抗氧化酶活性和RbohC、RbohF基因的表达量均明显降低,经DMTU、DPI和IMD预处理后再分别进行盐和干旱胁迫,H_2O_2积累量下降,但U0126预处理后再进行胁迫处理,H_2O_2积累量上升.说明NADPH氧化酶、MAP激酶级联途径、H_2O_2参与了盐、干旱胁迫下活性氧产生、抗氧化酶活性变化和RbohC、RbohF基因表达的调控.     

Erwinia carotovora ssp. carotovora Infection Induced ＂Defense Lignin＂ Accumulation and Lignin Biosynthetic Gene Expression in Chinese Cabbage （Brassica rapa L. ssp. pekinensis）

Erwinia carotovora subsp, carotovora （Ecc） infects and causes soft rot disease in hundreds of crop species including vegetables, flowers and fruits. Lignin biosynthesis has been implicated in defensive reactions to injury and pathogen infection in plants. In this work, variations of lignin content and gene expression in the molecular interaction between Chinese cabbage and Ecc were investigated. H2O2 accumulation and peroxidase activity were detected by 3, 3＇- Dimethoxybenzidine staining at mocked and Ecc-inoculated sites of Chinese cabbage leafstalks. Klason lignin content in inoculated plants increased by about 7.84%, 40.37%, and 43.13% more than that of the mocked site at 12, 24 and 72 h after inoculation, respectively. Gas chromatography detected more p-coumaryl （H） and less coniferyl （G） and sinapyl （S） monolignins in leafstalks of Chinese cabbage. All three monomers increased in Ecc-infected leafstalks, and the Ecc-induced ＂defense lignin＂ were composed of more G and H monolignins, and less S monolignin. After searching the expressed sequence tags （EST） data of Chinese cabbage, 12 genes putatively encoding enzymes involved in lignin biosynthesis were selected to study their expression. All of these genes could be induced by mock inoculation and Ecc infection, while the gene expression lasted for several more hours in the infected samples than in mocked and untreated plants. Our results indicated that ＂defense lignin＂ was different from the developmental lignin in composition; G and S monolignins were significantly induced in plants in response to the soft rot Ecc; thus, lignin biosynthesis was differentially regulated and played a role in plant response to the soft rot Ecc.     

Maladaptation beyond a geographic range limit driven by antagonistic and mutualistic biotic interactions across an abiotic gradient

Species’ geographic range limits often result from maladaptation to the novel environments beyond the range margin. However, we rarely know which aspects of the n‐dimensional environment are driving this maladaptation. Especially of interest is the influence of abiotic versus biotic factors in delimiting species’ distributions. We conducted a 2‐year reciprocal transplant experiment involving manipulations of the biotic environment to explore how spatiotemporal gradients in precipitation, fatal mammalian herbivory, and pollination affected lifetime fitness within and beyond the range of the California annual plant, Clarkia xantiana ssp. xantiana. In the first, drier year of the experiment, fitness outside the range edge was limited mainly by low precipitation, and there was some evidence for local adaptation within the range. In the second, wetter year, we did not observe abiotic limitations to plant fitness outside the range; instead biotic interactions, especially herbivory, limited fitness outside the range. Together, protection from herbivory and supplementation of pollen resulted in three‐ to sevenfold increases in lifetime fitness outside the range margin in the abiotically benign year. Overall, our work demonstrates the importance of biotic interactions, particularly as they interact with the abiotic environment, in determining fitness beyond geographic range boundaries.