Multilocus sequence typing analysis of Italian Xanthomonas campestris pv. campestris strains suggests the evolution of local endemic populations of the pathogen and does not correlate with race distribution

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot in Brassicaceae. It is widespread in Italy and severe outbreaks occur under conditions that favour disease development. In this study a multilocus sequence typing approach (MLST) based on the partial sequence of seven loci was applied to a selection of strains representative of the main areas of cultivation and hosts. The aim was to investigate whether the long tradition of brassica crops in Italy has influenced the evolution of different Xcc populations. All loci were polymorphic; 14 allelic profiles were identified of which 13 were unique to Italian strains. Based on the seven loci, the most common genotype within the Italian Xcc strains (AP1) was also the most representative genotype found in worldwide Xcc strains. This genotype was included in a new clonal complex in addition to three other clonal complexes already identified in Xcc populations. The phylogenetic reconstruction using a concatenated dataset of four conserved protein-coding genes, dnaK, fuyA, gyrB and rpoD, showed that the Italian strains belonged to two genetic groups. Physiological races were also investigated for the first time in Italy. The race structure of Xcc was determined by inoculating eight differential Brassica lines belonging to five species and showed that, in Italy, race 4 is the most widespread, followed by races 1 and 6. No correlation was found between allelic profiles, host of isolation, geographical origin and races, although a prevalent race was identified within the same clonal complex.     

Are road verges corridors for weed invasion? Insights from the fine‐scale spatial genetic structure of Raphanus raphanistrum

Raphanus raphanistrum (Brassicaceae) is considered amongst the world's worst agricultural weeds. We address critical issues in its management by studying the pathway of colonisation at local scales. For this, we assessed the small‐scale spatial genetic structure of 231 samples collected from three different sites across the Cape Floristic Region, South Africa, using 11 nuclear microsatellite markers. Although natural pollen and seed dispersal were expected to be restricted, we found no significant relationship between genetic and geographical distance within sites. Instead, our results suggest that R. raphanistrum had colonised new habitats via jump dispersal, rather than through natural diffusive dispersal at local scales. We did not find evidence for road verges as dispersal corridors, as evidenced by a lack of isolation‐by‐distance at local scales. Instead, the absence of spatial genetic structure suggests that R. raphanistrum had rapidly spread throughout its current range, possibly facilitated by human‐mediated actions. Management plans addressing containment or suppression of the weedy species R. raphanistrum (and possibly other weedy species) should take the high degree of connectivity between distant geographical localities into account.     

Suppression of clubroot (Plasmodiophora brassicae) development in Arabidopsis thaliana by the endophytic fungus Acremonium alternatum

This study investigated the ability of an endophytic fungus Acremonium alternatum to reduce clubroot formation in the model plant Arabidopsis thaliana, which is highly susceptible to Plasmodiophora brassicae . Quantitative PCR demonstrated that A. alternatum colonized the P. brassicae -infected roots and shoots of the host plant. When Arabidopsis plants were co-inoculated with P. brassicae and A. alternatum , gall formation was reduced as shown by the reduction of the disease index (DI) by up to 50% compared to plants only infected with P. brassicae, whereas the infection rate was lowered by about 20% only in several, but not all, experiments. Clubroot was similarly suppressed when plants were inoculated with autoclaved A. alternatum spores or spore extracts, showing that viable spores were not needed. However, A. alternatum spores did not inhibit P. brassicae resting spore germination. Compared to the normal root galls, the smaller root galls on A. alternatum -inoculated plants contained fewer resting spores of the clubroot pathogen. It was thus hypothesized that inoculation with A. alternatum delayed the development of P. brassicae . Using quantitative RT-PCR to monitor the expression of P. brassicae genes differentially expressed during the development of the disease, a delayed pathogen development was corroborated. Furthermore, greenhouse experiments identified a time window in which the endophyte had to be administered, where the latest effective time point was 5 days before inoculation with P. brassicae and the optimum treatment was to administer A. alternatum and P. brassicae at the same time. These results indicate that A. alternatum and perhaps similar endophytes could be useful for the management of clubroot disease.     

Germination of <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis> resting spores stimulated by a non-host plant

Plant-induced germination of Plasmodiophora brassicae resting spores was studied in a laboratory experiment. Spore reaction was analysed in nutrient solution with exudates from growing roots of different plant species – one host plant (Brassica rapa var. pekinensis) and four non-host plants (Lolium perenne, Allium porrum, Secale cereale and Trifolium pratense) – and in controls with distilled water and nutrient solution. It was found that root exudates from L. perenne stimulated spore germination more than exudates from the other plants, including those from the host plant. The effect could not be explained by differences in the nutritional composition of the solutions due to differential uptake of the plant species, or by differences in root activity, measured as exudation of soluble sugars. This is the first time such a separation of factors has been done in analysing the influence of plants on P. brassicae germination. Although stimulation of P. brassicae resting spore germination is not restricted to the presence of host plants, it seems to vary depending on the plant species.     

十字花科蔬菜害虫种类及综合治理技术

针对近年来江门市蔬菜种植过程中害虫种类繁多、化学农药使用量增加,且抗药性不断增强,防治工作困难的现状,进行了多年实践,在分析江门市十字花科蔬菜害虫发生种类及发生特点的基础上,提出通过采用清除残株落叶、合理轮作等农业防治措施,设置杀虫灯、黄板、性诱剂等物理防治措施,释放寄生蜂、捕食性天敌等生物防治措施,辅之以合理使用高效...     

Variation in floral scent compounds recognized by honeybees in Brassicaceae crop species

Floral scent attracts pollinators. We investigated the floral scent compounds recognized by pollinators in six Brassica crop species, including allogamous species with different genomes and autogamous species with two parental genomes and radish (Raphanus sativus). Biologically active compounds recognized by honeybees were screened from all floral compounds by combined gas chromatography–electroantennogram analysis and their profiles were determined by gas chromatography–mass spectrometry. Fourteen of the 52 compounds were active. All accessions had more than two active compounds, but the compounds greatly differed between the two genera. On the basis of similarities in whether active compounds were presence or absence, their amount and their composition ratio, we divided the Brassica accessions into three to five groups by cluster analyses. Most groups were composed of a mixture of allogamous and autogamous species sharing same genome, indicating that the variation depended on genome, not species. These results suggest that all species require pollinator visits for reproduction, despite their different reproductive systems. However, the inter-genus and intra-specific variations shown by the multiple groups within a species might cause different visitation frequencies by pollinators between genera and among accessions within a species, resulting in insufficient seed production in some accessions or species.     

十字花科植物PIN3基因调控元件及表达分析

为探明十字花科不同物种PIN3基因的表达调控差异,从拟南芥(Arabidopsis thaliala)、红花荠菜(Capsella rubella)、白菜(Brassica rapa)、油菜(Brassica napus)和甘蓝(Brassica oleracea)等17个已完成基因组测序的十字花科物种基因组数据库中鉴定出18个PIN3同源基因。对这18个同源基因编码序列上游1 500bp启动子区域顺式作用元件进行预测及比对分析,结果发现,光响应、多种激素响应及胁迫响应等相关元件呈现出较大差异,其中生长素以及向地性响应相关元件具有较高的保守性。克隆普通荠菜PIN3同源基因(CbPIN3)并构建普通荠菜PIN3基因启动子的GUS报告系统CbPIN3pro::GUS转化拟南芥。与拟南芥基因表达数据库比较,荠菜启动子表达与拟南芥PIN3具有基本一致的组织表达模式,但也存在一定的组织差异,说明植株形态建成中生长素极性转运调控的差异。拟南芥和荠菜PIN3的表达差异是十字花科物种适应性进化模式的缩影。     

Uptake and Translocation of Copper in Brassicaceae

Abstract

Ten cvs. of four Brassicaceae species were tested to evaluate their copper (Cu) uptake and translocation. Germination and root length tests indicated that Brassica juncea cv. Aurea and Raphanus sativus cvs. Rimbo and Saxa were the species with the highest germinability and longest roots at Cu concentrations ranging from 25 up to 200 µM. Raphanus sativus cv. Rimbo grown in hydroponic culture at increasing Cu concentrations (from 0.12 up to 40 µM) for 10 days produced a relatively high biomass (17.2 mg plant^?1) at the highest concentration and had a more efficient Cu translocation (17.8%) in comparison with cvs. Aurea and Saxa. The potential of cv. Rimbo for Cu uptake was then followed for 28 days at 5, 10, and 15 µM Cu. In comparison with the control, after 28 days of growth the 15 µM Cu‐treated plants showed a reduction in the tolerance index (?40%) and in the above‐ground dry biomass (?19%). On the contrary, an increase in the below‐ground dry weight was observed (+35%). Copper accumulated during the growth period both in the below‐ and above‐ground parts (about 14 and 4 µg plant^?1 at 10 and 15 µM Cu, respectively), but the translocation decreased from 50 to 30% in the last week at all the concentrations used. In addition, cv. Rimbo grown in a multiple element [cadmium (Cd), chromium (Cr), Cu, lead (Pb), and zinc (Zn)] naturally‐contaminated site accumulated all elements in the above‐ground part in a range from 5 to 62 µg plant^?1.     

Incidence of cabbage maggot (Diptera: Anthomyiidae) infestation and plant damage in seeded Brassica fields in California's central coast

The temporal incidence of cabbage maggot, Delia radicum L. was investigated using three exclusion cage experiments, one each during spring, summer and fall, in broccoli fields as well as two surveys, one each in three broccoli fields, and sixteen turnip plantings in central coast of California. In the cage experiments, sets of broccoli plants were exposed to natural populations of D. radicum flies for ∼14-d periods after plant emergence throughout the growing season. For the surveys, soil, root samples, and yellow sticky traps were collected every week from broccoli fields to determine number of eggs, maggots, feeding-injury and adults. Only roots were sampled from turnip plantings to determine feeding-injury. In all three cage experiments, feeding injury from D. radicum maggot was less during the first 14-d than ∼15–28 d after plant emergence (DAE). In the summer and fall, feeding injury by D. radicum was less during 29–42 and 43–56 DAE than it was 15–28 DAE. In the survey of broccoli fields, a greater number of D. radicum eggs were detected starting the fourth week after planting (WAP). Similarly, an increase in number of D. radicum maggots and feeding injury was observed at fifth and sixth WAP, respectively. However, adults were abundant throughout the growing period. In the turnip survey, increase in injury from D. radicum feeding did not appear until the fifth WAP. Overall, these studies indicate that increased incidence of D. radicum was delayed by about two to three weeks after plant emergence. The implications of these results for timing of insecticide application for D. radicum in the central coast of California are discussed.     

Insecticides applied to soil of transplant plugs for Bagrada hilaris (Burmeister) (Hemiptera: Pentatomidae) management in broccoli

Chemical control with insecticides, typically applied as foliar sprays or chemigation, is the primary tactic used to manage Bagrada hilaris (Burmeister) (Hemiptera: Pentatomidae). We evaluated the efficacy of 14 insecticides, including both systemic and non-systemic insecticides, against B. hilaris applied as a seedling tray drench. Experiments were conducted in both greenhouse and field settings. In all experiments, we used the maximum label rate of insecticides and calculated dose per seedling based on this rate and standard plant density per hectare. Each seedling in the tray received 2-mL insecticide solution, and the seedlings were then exposed to B. hilaris adults after transplanting in cages for greenhouse experiments or natural B. hilaris populations in the field experiments. A scale system (0–4) was used to evaluate the severity of B. hilaris feeding injury on leaves where 0 = no injury and 4 = >75% of the leaf margins with B. hilaris feeding injury. We evaluated damage using the rating system, percentage of damaged leaves, number of feeding injury sites, as well as plant height, leaf width, and fresh and dry weight. In the greenhouse experiment, percentage of injured leaves, number of injury sites, and damage rating were significantly lower for transplants treated with acetamiprid, clothianidin, dinotefuran, imidacloprid, and thiamethoxam, and cyclaniliprole compared with the untreated. There was a relationship between feeding injury sites and plant height, leaf width, fresh and dry weight (R² > 0.5) in both 2015 field experiments. Similarly, the transplants treated with acetamiprid, clothianidin, dinotefuran, imidacloprid, thiamethoxam, thiamethoxam + chlorantraniliprole, imidacloprid + β-cyfluthrin and cyclaniliprole had significantly lower damage ratings than those treated with chlorpyrifos, bifenthrin, tolfenpyrad, flonicamid, cyantraniliprole, spinetoram and the untreated control. For transplanted broccoli, treatment of transplant plugs with neonicotinoid insecticides prior to planting can be an effective method for controlling B. hilaris.