What proteomic analysis of the apoplast tells us about plant–pathogen interactions

Plant pathogens have developed different strategies during their evolution to infect and colonize their hosts. In the same way, plants have evolved different mechanisms acting against potential pathogens trying to infect and colonize their tissues. Regulation of a wide variety of proteins is required in order to perceive the pathogen and to activate the plant defence mechanisms. The apoplast is the first compartment where these recognition phenomena occur in most plant–pathogen interactions, allowing the exchange of different molecules and facilitating inter‐ and intracellular communication in plant cells. Proteomic analysis of the apoplast in recent years has found the initial biochemical responses involved in pathogen recognition and early defence responses. However, this proteomic approach requires some specific experimental conditions to obtain an extract free of cytoplasmic proteins and nonprotein contaminants that affect the subsequent stages of separation and quantification. Obtaining the highest proportion of proteins from the apoplastic space in infected tissues requires different steps such as extraction of apoplastic washing fluids and preparation of total secreted proteins (protein precipitation, solubilization, separation and digestion). Protein identification using mass spectrometry techniques and bioinformatics tools identifying peptides for the extracellular exportation is required to confirm the apoplastic location. This review compiles the most commonly used techniques for proteomic studies, focusing on the early biochemical changes occurring in the apoplast of plants infected by a wide range of pathogens. The scope of this approach to discover the molecular mechanisms involved in the plant–pathogen interaction is discussed.     

Seed Shattering of Cañahua (Chenopodium pallidicaule Aellen)

Cañahua (Chenopodium pallidicaule Aellen) is a semi‐domesticated relative of quinoa (Chenopodium quinoa Willd.) with high nutritious quality. It is tolerant to frost, drought, saline soils and pests. One seed yield limitation is seed loss during the maturity stages. Two greenhouse experiments in Denmark and field experiments in Bolivia were carried out to determine seed shattering in landraces and cultivars with different growth habits. 15–21 % of the seed shattering in the fields took place whilst the plants still were flowering and 25–35 % during physiological maturity. Seed shattering varied between locations on the Bolivian Altiplano. Cañahua types with the semi‐prostrate growth (‘lasta’) had the highest seed shattering rate in the greenhouse experiments. The Umacutama landrace had lower seed shattering (1 %) than the cultivar Kullaca (7.2 %) both of the ‘lasta’ type. Under field conditions, the cultivar Illimani with the erect growth (‘saihua’) had the highest seed shattering rate (6.4–33.7 %) at both locations and at four different sowing dates. The Umacutama had the lowest rate (0.5–1.5 %). There were no significant differences between plants of the ‘lasta’ and the ‘saihua’ types. The landrace had significantly less seed loss than the cultivars. However, in the greenhouse, the landrace yield was approximately 25 % lower than the yields of the cultivars. In general, cañahua cultivars had higher yield compared to landraces, but also a higher seed shattering rate. Landraces may be used in breeding programmes to develop high‐yielding cultivars with reduced seed shattering.     

Some Agronomic Strategies for Organic Quinoa (Chenopodium quinoa Willd.)

Quinoa is a potential new seed crop for protein feed and human consumption in Europe, with tolerance to a range of abiotic stresses. For this purpose the study was planned to analyse the effect of important agronomic strategies like nitrogen level, N application strategy, row spacing and harvest time on yield and quality of quinoa. The experiments took place in the field of the experimental station of the Faculty of Science, University of Copenhagen. Three levels of organic nitrogen from slurry was used (60, 120 and 180 kg N ha^?1), supplied either all at once at sowing, or split between sowing and beginning of the reproductive phase. The effect of row spacing and harvest time was studied by harvesting seeds at seed maturity, which occurred 2–3 weeks prior to the mechanical harvest by threshing, and a couple of months after. Yield increased significantly (P ≤ 0.05) with an application up to 180 kg N ha^?1, reaching 2200 kg ha^?1. Increasing N also caused a significantly increased seed weight (up to 3.3 mg) and protein content (up to 17 %). N level did not affect number and amount of weeds. Split application with part of the N applied at bud formation did not have a significant effect on yield. Delayed harvest had a negative influence on seed weight, whereas protein content was stable after harvesting even a month after seed maturity. A late harvest significantly reduced seed germination, being reduced by 50 % after a 2‐month delay. A conclusion from this study is that both yield and protein content of seed can be manipulated by N level and application strategy. Harvest time is important for securing a high seed quality measured as seed germination, seed weight and protein content. A fast germination of quinoa is an important characteristic demonstrating that the crop has good possibilities for being well‐established in the field when free from weeds at the time of sowing. The choice of row spacing is important and depends on weed control method. Weed control strategy should be developed based on modern precision tools.     

Antifungal properties of terpenoids in Picea abies against Heterobasidion parviporum

The antifungal properties of 14 major oleoresin‐constitutive terpenoids of Norway spruce (Picea abies) against Heterobasidion parviporum were evaluated in vitro at three gradient concentrations, 0.1, 0.2 and 0.4 μmol/cm², on potato dextrose agar medium. Eight monoterpene hydrocarbons (+)‐ and (?)‐α‐pinene, (?)‐β‐pinene, (+)‐3‐carene, myrcene, (+)‐ and (?)‐limonene, terpinolene; four oxygenated monoterpenes α‐terpineol, terpinen‐4‐ol, 1,8‐cineole, bornyl acetate; and two resin acids abietic acid and dehydroabietic acid were selected. Abietic and dehydroabietic acids showed the highest antifungal activities at all concentrations. Among oxygenated monoterpenes, bornyl acetate and α‐terpineol showed antifungal activity at the highest concentration. Among monoterpene hydrocarbons, (+)‐α‐pinene showed similar activity to terpinen‐4‐ol and 1,8‐cineole at the highest concentration; however, it was lower than α‐terpineol and bornyl acetate. Other monoterpene hydrocarbons inhibited mycelial growth by <10%. Re‐extraction of medium surfaces after the test period revealed that most α‐terpineol and terpinen‐4‐ol remained unchanged on the surface but monoterpene hydrocarbons completely disappeared. However, notable fungal transformed products were observed on surfaces applied with 1,8‐cineole and bornyl acetate. Thus, mycelial growth inhibition of monoterpenoids might be caused by complex functions such as biodegradation and/or detoxification by interaction between mycelium and compound. These results provide a basis for future studies considering the role of chemodiversity in the comprehensive chemical defence of P. abies.     

Effects of Land‐Related Policies on Land Development during a Real Estate Boom and a Recession

Our research evaluates and compares the effects of three land‐related policies on land development in a sprawling metropolitan area during a real estate boom and a recession. Our findings suggest that during a real estate boom 1) the urban growth boundary (UGB) serves its purpose of attracting urban development inside the given boundary during a boom while its effectiveness diminishes with increased development pressure from lower‐valued land outside of the UGB during a recession period, 2) the agricultural zone works well for restraining new development during a boom period while agriculture zoned parcels are more likely developed during a recession period, and 3) an increase in the land‐value tax bill increases the incentive for development during a recession period while higher tax bills do not affect development during a boom period. In anticipation of a re‐emergence of urban sprawl with the recovery of the real estate market, our findings imply that land planners and others concerned with sprawling development should pay more attention to development in sprawl‐prone areas during recession periods.