This was the attached instructions from the email.
Extra Credit Assignment: Due end of day November 25th.
Write a synopsis of the 3 papers.
1.Choudhury & McRoberts 2018. (Dr. Choudhury is a professor at UTRGV)
2.Wei et al 2019 (Dr. Cristoffersen is a professor at UTRGV).
3.Martinez et al 2020 (all authors are students or faculty at UTRGV).
For each write only two paragraphs. Paragraph one should include the reason for the research, the importance of the research, the scientific question being asked by the scientists and the hypothesis/hypotheses being tested. Paragraph two should include whether the results supported or did not support the research, how the results supported or did not support the research, how the results add to the field of study and/or what research still needs to be done. Write in simple words. Straight to the point.
Every two paragraphs are about 1.7 potential points that can be earned by student.
If paper synopsis seems generally solid the student may earn 5 ppts. max to final average. (*hint* final will be open ended)
Cheating will result in a zero for the extra credit points. No quotes may be used in the essay; put everything into your own words. You can use sources other than the paper to help write the essay.
Study up class!
Temperature and light effects on in vitro germination of Peronospora effusa sporangia SHORT COMMUNICATION Temperature and light effects on in vitro germination of Peronospora effusa sporangia Robin Alan Choudhury1,2 & Neil McRoberts1 Received: 25 August 2017 /Accepted: 7 December 2017 /Published online: 22 December 2017 # Sociedade Brasileira de Fitopatologia 2017 Abstract Spinach downy mildew, caused by the biotrophic oomycete Peronospora effusa, is an economically important disease that is found in all spinach growing regions of the US. To effectively predict disease risk we need to understand the response of P. effusa to different environmental conditions. We conducted several in vitro germination assays, exposing P. effusa sporangia to different temperature and lighting conditions. Between 5 and 25 °C under constant darkness, germination of P. effusa sporangia on water agar declined with increasing temperatures. These results were qualitatively different from a previous study of P. effusa germi- nation that found a bimodal response curve, with increased germination at lower and higher temperatures. Time course studies revealed that sporangia consistently germinated within the first 12 h of plating, regardless of incubation temperature. Blue light exposure significantly reduced sporangial germination when compared with sporangia exposed to red, yellow, or no light. Light intensity and color significantly impacted germination, although the effect of color varied by light intensity. Keywords Environmental effects . Germination . Light . Peronospora effusa . Spinach downymildew . Spinacia oleracea Spinach downy mildew, caused by the biotrophic oomycete Peronospora effusa (Grev.) Rabenh (formerly P. farinosa f. sp. spinaciae) (Choi et al. 2007), is the most important threat to spinach productionworldwide (Correll et al. 2011). Spinach downy mildew, like many other downy mildew diseases, typ- ically occurs in cool, moist environments (Choudhury et al. 2016). Coastal California frequently has cool morning tem- peratures and thick layers of fog and low clouds (Choudhury et al. 2016) and produces approximately 40% of the annual US fresh market crop of spinach (USDA NASS 2016). While these cool temperatures can persist well into the afternoon for some portions of the region, more southern areas of coastal California frequently have warm, dry afternoon conditions for much of the year. Outbreaks of spinach downy mildew fre- quently occur despite these warmer and drier conditions, and can lead to large crop failures (Choudhury et al. 2016). These disease outbreaks in warmer regions have many growers con- cerned that P. effusa may display phenotypic plasticity, and have a wider range of optimal environmental conditions for growth compared with other downy mildew pathogens. To effectively predict disease risk we need to understand the response ofP. effusa to different environmental conditions. However, compared with many other downy mildew patho- gens, relatively little research has been done on the optimal environmental conditions for growth and sporulation of P. effusa. Recent field studies have revealed that environmen- tal variables play a large role in the epidemiology of this dis- ease (Choudhury et al. 2016). However, without understand- ing how individual environmental factors impact different stages of the pathogen life cycle, it is difficult to create an accurate risk prediction system. These controlled environment studies help to clarify how environmental variables affect the pathogens growth, reproduction and spread, without other confounding environmental effects. An early report of germination rates of a P. effusa isolate fromCalifornia suggested that while P. effusa could germinate between 0 and 30 °C, the optimal temperatures for germina- tion were between 10 and 15 °C (Cook 1937). A later study from the Netherlands suggested that spinach downy mildew isolates had a bimodal reaction to increasing temperatures, and Section Editor: Sarah J. Pethybridge * Robin Alan Choudhury
[email protected] 1 Department of Plant Pathology, University of California, Davis, Davis, CA 95616, USA 2 Present address: Plant Pathology Department, University of Florida, Gainesville, FL 32601, USA Tropical Plant Pathology (2018) 43:572–576 https://doi.org/10.1007/s40858-017-0204-y http://crossmark.crossref.org/dialog/?doi=10.1007/s40858-017-0204-y&domain=pdf mailto:
[email protected] that P. effusa sporangia germinated between the temperatures 0 and 30 °C (Frinking et al. 1981). Interestingly, they found sporangial germination peaked at both 10 and 25 °C, with a reduction of germination around 20 °C. A bimodal tempera- ture response in germination could help to explain how dis- ease outbreaks can occur in warm regions as well as cooler regions through local adaptation of the pathogen. While understanding the overall range of optimal temper- atures is important for future disease prediction efforts, under- standing the timescale of sporangial germination is critical to the timing of protective fungicide sprays (Caffi et al. 2010). Most non-systemic fungicides, like dithiocarbamates and chloroisophthalonitriles, provide protective activity rather than curative or eradicative activity (Gisi and Sierotzki 2008). These fungicides provide protection by inhibiting the growth of the pathogen before it has entered the plant. Correct timing of fungicide applications can help to prevent the estab- lishment of fungal pathogens in host plants. Timing the fun- gicide application is critical to ensure that the susceptible ger- mination tube is exposed to prevent infection and crop failure. In addition to temperature and relative humidity, the quality and quantity of light can also have dramatic effects on growth and sporulation of downy mildew pathogens. The overall quantity and period of light can regulate different processes of oomycete biology, including sporulation and infection (Cohen et al. 2013;Wang et al. 2011). In addition to the effects from the total amount of light exposure, many oomycetes are affected by specific spectra of light. Some oomycetes are inhibited by blue light, which can reduce sporulation and growth (Cohen et al. 1975; Cohen 1976; Cohen and Eyal 1977). Interestingly, a recent report suggests that the basil downy mildew pathogen P. belbahrii is most negatively af- fected by red light, which strongly suppresses sporulation (Cohen et al. 2013). Some basil growers have taken advantage of this detrimental effect on sporulation by using red light in susceptible basil greenhouses, thereby reducing downy mil- dew epidemic severity (Cohen et al. 2013). Understanding how P. effusa is affected by different qualities and quantities of light may help to guide future control strategies for spinach downy mildew disease. In this study, we exposed P. effusa sporangia to different tem- perature and lighting conditions and quantified detrimental effects on in vitro germination. The goals of this study were to understand: (1) how California isolates of P. effusa are affected by temperature; (2) how temperature affects the time to germi- nation of sporangia; and (3) how different qualities and quantities of light affect sporangial germination. Six isolates of P. effusawere collected from diseased fields in the Salinas Valley in 2015 and 2016 (Table 1). Isolates were considered distinct if they were localized as small foci (<20 cm)="" that="" were="" spatially="" separated="" from="" other="" outbreaks="" (="">2 m), therefore likely arising from the expansion of a single infection event. Approximately 20 diseased leaves from each isolate were collected into sterile bag containers, and two bags were collected from each outbreak source. Bags were sealed and stored at 4 °C in a cooler until processing (<6 h). sporangia were removed from infected leaves by applying 50 ml of chilled sterile distilled water into the sealed bags and shaking vigorously for five minutes. the resulting spore suspension was standardized to 1 × 105 spores/ml using a haemocytometer. spore suspensions were maintained on ice while being prepared for experimental use. to test the effects of temperature of germination, 150 μl of p. effusa 105 sporangia/ml spore suspension were plated and spread uniformly on 2% water agar on 100 × 15 mm plates. plates were immediately stored in crisper containers, square transparent plastic boxes with lids, and the containers were placed into incubation chambers (model e7, conviron ltd.) maintained at 5, 10, 15, 20, and 25 °c in darkness. incubation chamber temperatures were confirmed using analog thermom- eters. ten plates were used for each temperature treatment and isolate (table 1). at 24 h after plating, crisper boxes were removed from incubation chambers. plates were examined under a 10× light microscope for sporangial germination. one hundred sporangia from each plate were rated for germi- nation status. sporangia that had visibly developed sub- terminal or lateral germ tubes were considered germinated. experiments were repeated twice. the effect of temperature on sporangial germination over time was measured for p. effusa isolate dnm003. plates were treated as described above, and sporangial germination was measured at 4, 8, 12, and 24 h after plating. the experiment was repeated twice. to estimate the effect of light quality on p. effusa sporan- gial germination, the lids of crisper boxes were fitted with red, table 1 information on the source of peronospora effusa isolates used in this study isolate number variety location collection month temperaturea (°c) dnm001 violin king city, ca, usa june, 2015 18.1 dnm002 platypus hollister, ca, usa july, 2015 19.5 dnm003 carmel hollister, ca, usa july, 2015 19.5 dnm004 emilia king city, ca, usa may, 2016 15.5 dnm005 emilia king city, ca, usa may, 2016 15.5 dnm006 viroflay watsonville, ca, usa july, 2016 13.7 a average temperature(°c) during the month of collection of isolate source location trop. plant pathol. (2018) 43:572–576 573 yellow, green, and blue color correction gels (cowboy studio llc.) to allow exposure to specific qualities of light. the quality and quantity of light was measured within the sealed crisper boxes using a spectrometer (black comet model, stellarnet) using the spectrawiz software (stellarnet). the wavelength spectra of the crisper boxes used with color cor- rection gels shows non-specific wavelengths outside of the target spectra (fig. 1). the bottoms and sides of the crispers were covered in aluminum foil to prevent ambient light. the crisper box used to test the effect of constant darkness on germination was completely covered with aluminum foil, and the crisper box used to test no filter was left unaltered. seventy-five microliters of p. effusa isolate dnm003 spore suspension (105 sporangia/ml) were plated and spread uni- formly on 2% water agar on 60 × 15 mm plates. ten plates were laid in a single layer on the bottom of the crisper boxes, and boxes were immediately moved into a growth chamber. the effects of light intensity and temperature were evaluated alongside light quality in a fully factorial design. the growth chamber was set to either constant full fluorescent and incan- descent lamps (full light, fig. 1) or constant half-power fluo- rescent lamps (low light, fig. 1), at either 4 or 18 °c. these temperatures were chosen to reflect the average low and high temperatures experienced during the growing season in coast- al california. the plates were removed and measured for spo- rangial germination as described above at 24 h after plating. all experiments were repeated twice. means comparisons between different treatment tempera- tures, time periods, and lighting conditions were completed using fisher’s least significant difference test, implemented h).="" sporangia="" were="" removed="" from="" infected="" leaves="" by="" applying="" 50="" ml="" of="" chilled="" sterile="" distilled="" water="" into="" the="" sealed="" bags="" and="" shaking="" vigorously="" for="" five="" minutes.="" the="" resulting="" spore="" suspension="" was="" standardized="" to="" 1="" ×="" 105="" spores/ml="" using="" a="" haemocytometer.="" spore="" suspensions="" were="" maintained="" on="" ice="" while="" being="" prepared="" for="" experimental="" use.="" to="" test="" the="" effects="" of="" temperature="" of="" germination,="" 150="" μl="" of="" p.="" effusa="" 105="" sporangia/ml="" spore="" suspension="" were="" plated="" and="" spread="" uniformly="" on="" 2%="" water="" agar="" on="" 100="" ×="" 15="" mm="" plates.="" plates="" were="" immediately="" stored="" in="" crisper="" containers,="" square="" transparent="" plastic="" boxes="" with="" lids,="" and="" the="" containers="" were="" placed="" into="" incubation="" chambers="" (model="" e7,="" conviron="" ltd.)="" maintained="" at="" 5,="" 10,="" 15,="" 20,="" and="" 25="" °c="" in="" darkness.="" incubation="" chamber="" temperatures="" were="" confirmed="" using="" analog="" thermom-="" eters.="" ten="" plates="" were="" used="" for="" each="" temperature="" treatment="" and="" isolate="" (table="" 1).="" at="" 24="" h="" after="" plating,="" crisper="" boxes="" were="" removed="" from="" incubation="" chambers.="" plates="" were="" examined="" under="" a="" 10×="" light="" microscope="" for="" sporangial="" germination.="" one="" hundred="" sporangia="" from="" each="" plate="" were="" rated="" for="" germi-="" nation="" status.="" sporangia="" that="" had="" visibly="" developed="" sub-="" terminal="" or="" lateral="" germ="" tubes="" were="" considered="" germinated.="" experiments="" were="" repeated="" twice.="" the="" effect="" of="" temperature="" on="" sporangial="" germination="" over="" time="" was="" measured="" for="" p.="" effusa="" isolate="" dnm003.="" plates="" were="" treated="" as="" described="" above,="" and="" sporangial="" germination="" was="" measured="" at="" 4,="" 8,="" 12,="" and="" 24="" h="" after="" plating.="" the="" experiment="" was="" repeated="" twice.="" to="" estimate="" the="" effect="" of="" light="" quality="" on="" p.="" effusa="" sporan-="" gial="" germination,="" the="" lids="" of="" crisper="" boxes="" were="" fitted="" with="" red,="" table="" 1="" information="" on="" the="" source="" of="" peronospora="" effusa="" isolates="" used="" in="" this="" study="" isolate="" number="" variety="" location="" collection="" month="" temperaturea="" (°c)="" dnm001="" violin="" king="" city,="" ca,="" usa="" june,="" 2015="" 18.1="" dnm002="" platypus="" hollister,="" ca,="" usa="" july,="" 2015="" 19.5="" dnm003="" carmel="" hollister,="" ca,="" usa="" july,="" 2015="" 19.5="" dnm004="" emilia="" king="" city,="" ca,="" usa="" may,="" 2016="" 15.5="" dnm005="" emilia="" king="" city,="" ca,="" usa="" may,="" 2016="" 15.5="" dnm006="" viroflay="" watsonville,="" ca,="" usa="" july,="" 2016="" 13.7="" a="" average="" temperature(°c)="" during="" the="" month="" of="" collection="" of="" isolate="" source="" location="" trop.="" plant="" pathol.="" (2018)="" 43:572–576="" 573="" yellow,="" green,="" and="" blue="" color="" correction="" gels="" (cowboy="" studio="" llc.)="" to="" allow="" exposure="" to="" specific="" qualities="" of="" light.="" the="" quality="" and="" quantity="" of="" light="" was="" measured="" within="" the="" sealed="" crisper="" boxes="" using="" a="" spectrometer="" (black="" comet="" model,="" stellarnet)="" using="" the="" spectrawiz="" software="" (stellarnet).="" the="" wavelength="" spectra="" of="" the="" crisper="" boxes="" used="" with="" color="" cor-="" rection="" gels="" shows="" non-specific="" wavelengths="" outside="" of="" the="" target="" spectra="" (fig.="" 1).="" the="" bottoms="" and="" sides="" of="" the="" crispers="" were="" covered="" in="" aluminum="" foil="" to="" prevent="" ambient="" light.="" the="" crisper="" box="" used="" to="" test="" the="" effect="" of="" constant="" darkness="" on="" germination="" was="" completely="" covered="" with="" aluminum="" foil,="" and="" the="" crisper="" box="" used="" to="" test="" no="" filter="" was="" left="" unaltered.="" seventy-five="" microliters="" of="" p.="" effusa="" isolate="" dnm003="" spore="" suspension="" (105="" sporangia/ml)="" were="" plated="" and="" spread="" uni-="" formly="" on="" 2%="" water="" agar="" on="" 60="" ×="" 15="" mm="" plates.="" ten="" plates="" were="" laid="" in="" a="" single="" layer="" on="" the="" bottom="" of="" the="" crisper="" boxes,="" and="" boxes="" were="" immediately="" moved="" into="" a="" growth="" chamber.="" the="" effects="" of="" light="" intensity="" and="" temperature="" were="" evaluated="" alongside="" light="" quality="" in="" a="" fully="" factorial="" design.="" the="" growth="" chamber="" was="" set="" to="" either="" constant="" full="" fluorescent="" and="" incan-="" descent="" lamps="" (full="" light,="" fig.="" 1)="" or="" constant="" half-power="" fluo-="" rescent="" lamps="" (low="" light,="" fig.="" 1),="" at="" either="" 4="" or="" 18="" °c.="" these="" temperatures="" were="" chosen="" to="" reflect="" the="" average="" low="" and="" high="" temperatures="" experienced="" during="" the="" growing="" season="" in="" coast-="" al="" california.="" the="" plates="" were="" removed="" and="" measured="" for="" spo-="" rangial="" germination="" as="" described="" above="" at="" 24="" h="" after="" plating.="" all="" experiments="" were="" repeated="" twice.="" means="" comparisons="" between="" different="" treatment="" tempera-="" tures,="" time="" periods,="" and="" lighting="" conditions="" were="" completed="" using="" fisher’s="" least="" significant="" difference="" test,="">6 h). sporangia were removed from infected leaves by applying 50 ml of chilled sterile distilled water into the sealed bags and shaking vigorously for five minutes. the resulting spore suspension was standardized to 1 × 105 spores/ml using a haemocytometer. spore suspensions were maintained on ice while being prepared for experimental use. to test the effects of temperature of germination, 150 μl of p. effusa 105 sporangia/ml spore suspension were plated and spread uniformly on 2% water agar on 100 × 15 mm plates. plates were immediately stored in crisper containers, square transparent plastic boxes with lids, and the containers were placed into incubation chambers (model e7, conviron ltd.) maintained at 5, 10, 15, 20, and 25 °c in darkness. incubation chamber temperatures were confirmed using analog thermom- eters. ten plates were used for each temperature treatment and isolate (table 1). at 24 h after plating, crisper boxes were removed from incubation chambers. plates were examined under a 10× light microscope for sporangial germination. one hundred sporangia from each plate were rated for germi- nation status. sporangia that had visibly developed sub- terminal or lateral germ tubes were considered germinated. experiments were repeated twice. the effect of temperature on sporangial germination over time was measured for p. effusa isolate dnm003. plates were treated as described above, and sporangial germination was measured at 4, 8, 12, and 24 h after plating. the experiment was repeated twice. to estimate the effect of light quality on p. effusa sporan- gial germination, the lids of crisper boxes were fitted with red, table 1 information on the source of peronospora effusa isolates used in this study isolate number variety location collection month temperaturea (°c) dnm001 violin king city, ca, usa june, 2015 18.1 dnm002 platypus hollister, ca, usa july, 2015 19.5 dnm003 carmel hollister, ca, usa july, 2015 19.5 dnm004 emilia king city, ca, usa may, 2016 15.5 dnm005 emilia king city, ca, usa may, 2016 15.5 dnm006 viroflay watsonville, ca, usa july, 2016 13.7 a average temperature(°c) during the month of collection of isolate source location trop. plant pathol. (2018) 43:572–576 573 yellow, green, and blue color correction gels (cowboy studio llc.) to allow exposure to specific qualities of light. the quality and quantity of light was measured within the sealed crisper boxes using a spectrometer (black comet model, stellarnet) using the spectrawiz software (stellarnet). the wavelength spectra of the crisper boxes used with color cor- rection gels shows non-specific wavelengths outside of the target spectra (fig. 1). the bottoms and sides of the crispers were covered in aluminum foil to prevent ambient light. the crisper box used to test the effect of constant darkness on germination was completely covered with aluminum foil, and the crisper box used to test no filter was left unaltered. seventy-five microliters of p. effusa isolate dnm003 spore suspension (105 sporangia/ml) were plated and spread uni- formly on 2% water agar on 60 × 15 mm plates. ten plates were laid in a single layer on the bottom of the crisper boxes, and boxes were immediately moved into a growth chamber. the effects of light intensity and temperature were evaluated alongside light quality in a fully factorial design. the growth chamber was set to either constant full fluorescent and incan- descent lamps (full light, fig. 1) or constant half-power fluo- rescent lamps (low light, fig. 1), at either 4 or 18 °c. these temperatures were chosen to reflect the average low and high temperatures experienced during the growing season in coast- al california. the plates were removed and measured for spo- rangial germination as described above at 24 h after plating. all experiments were repeated twice. means comparisons between different treatment tempera- tures, time periods, and lighting conditions were completed using fisher’s least significant difference test, implemented>20>