HORTSCIENCE 57(11):1436–1446. 2022. https://doi.org/10.

21273/HORTSCI16777-22 increased pressure on table beet and Swiss

chard seed and vegetable producers to maintain
foliar health. BLS was first found in sugar beet
Screening Table Beet and Swiss Chard crops in Utah and California in the first decade
of the 20th century and again in California in
for Resistance to Pseudomonas syringae 1913 (Brown and Jamieson, 1913). This was
followed by documentation of the disease
pathovar aptata in sugar beet crops in the Pacific Northwest
(Oregon and Northern California) in 1944
Emilee Gaulke and Irwin L. Goldman (Carsner, 1944). More recently, BLS has
Department of Horticulture, University of Wisconsin-Madison, 1575 Linden been reported on sugar beet in Georgia in
Drive, Madison, WI 53706 2012 (Dutta et al., 2014) and in Oregon in
2015 (Arabiat et al., 2016). The disease was
Keywords. AUDPC, bacterial leaf spot, disease resistance, Pseudomonas syringae, table first reported in Swiss chard in California be-
beet tween 1999 and 2003 (Koike et al., 2003). BLS
is continuing to spread to new geographic loca-
Abstract. Bacterial leaf spot (BLS) has emerged in the last few decades as an economi- tions, with a recent report in Arizona in 2019
cally important disease of both table beet (Beta vulgaris ssp. vulgaris) and Swiss chard (Nampijja et al., 2021). The first report in table
(Beta vulgaris ssp. cicla). BLS is caused by Pseudomonas syringae pv. aptata, which is beet in Ohio was in 2017 (Rotondo et al.,
spread readily on infected seeds. Symptoms appear as circular to irregular shaped, with 2020), although BLS had been documented in
a tan to dark brown center and a very dark border. Disease incidence and severity is de- table beet and Swiss chard seed crops in the Pa-
pendent on cool, humid conditions and can vary widely year to year depending on the en- cific Northwest for several years prior (Derie
vironment. Both the vegetative and reproductive phases of these biennial crops are et al., 2016; Safni et al., 2016). Jacobsen
susceptible to the pathogen. Table beet and Swiss chard commercial cultivars (n 5 21), (2009) noted that BLS has been observed on
table beet breeding lines (n 5 5), and table beet plant introductions (PIs) (n 5 26) were sugar beet and table beet in most production
screened for response to spray inoculation with P. syringae pv. aptata in a controlled areas of the United States as well as in Japan,
greenhouse setting. Plants were rated for severity of symptoms using percent of the area western European countries, Australia, New
of each pair of leaves (leaf set) with symptoms and an overall plant score assigned based Zealand, and Russia; and in Swiss chard crops
on the scores for each leaf pair. Accessions varied in BLS susceptibility. PI accessions in the United States as well as European coun-
were most variable, with the area under the disease progress curve (AUDPC) ranging tries, Asia, and Australia. There are currently
from 1.33 to 8.75. Highly significant differences among PIs were detected for disease no reports of resistance to BLS in either table
scores in the vegetative stage, beginning 21 days after inoculation. Screens during the re- beet or Swiss chard in either the vegetative or
productive growth stage showed the least variation in AUDPC among PIs. Although cul- reproductive phases of their life cycle.
tivars varied less than PIs, good BLS resistance (low disease scores) was noted for Symptoms of BLS include lesions, each
‘Touchstone Gold’, ‘Kestrel’, ‘Bull’s Blood’, ‘Rainbow’ chard, as well as PIs 222234 and with a tan to brown centers surrounded by a
NSL 28026. Accessions W451C, Red Cloud, Detroit Dark Red, and NSL 28020 were dark border, that are circular to irregular in
highly susceptible. There was no consistent association between disease score in the vege- shape (Khan et al., 2018; Nikolic et al., 2018).
tative and reproductive phases, suggesting that breeders may need to screen for BLS in Bacteria enter leaves through stomata, hydath-
both phases of the biennial life cycle. The more resistant PIs or cultivars identified in this odes, or injuries caused by insects, wind, hail,
study can be used in future efforts to breed for host resistance to BLS and to establish or mechanical damage, and thus, lesions can
mapping populations to better understand the genetic control of resistance, to aid in occur both along the leaf margin and across
breeding efforts. the leaf blade (Khan et al., 2018; Lamichhane
et al., 2015). As the disease progresses, lesions
merge to form large necrotic regions, and the
Table beet (Beta vulgaris ssp. vulgaris) growth, particularly in New York, in large
leaf can become deformed (Khan et al., 2018;
and Swiss chard (Beta vulgaris ssp. cicla) are part due to increased recognition and interest
specialty crops grown for fresh markets, proc- Nikolic et al., 2018).
in the health benefits of consuming beets
BLS is caused by the pathogen Pseudomo-
essing, and baby leaf salad greens. Wisconsin (Pethybridge et al., 2018).
nas syringae pv. aptata (PSA). Pseudomonas
and New York are the leading producers of ta- Table beet production quality and yield
syringae is a bacterial species complex that in-
ble beet in the United States with 1650 ha and can be affected by a wide variety of patho-
cludes strains of pathogens that affect most eco-
1280 ha harvested, respectively, in 2017 [U.S. gens at several stages, both pre- and postharv- nomically important crops, including monocots
Department of Agriculture (USDA), 2019]. est (Pethybridge et al., 2018). According to and herbaceous and woody dicots (Lamichhane
The table beet industry currently is experiencing Pethybridge et al. (2018), an important factor et al., 2015). The P. syringae species complex
in table beet quality and yield is foliar health. comprises seven phylogroups that can be
Foliar health is key for optimal photosynthe- differentiated by multilocus sequence analysis
Received for publication 8 Jul 2022. Accepted sis, but also because beets grown for the fo- or whole genome sequence analysis (Gomila
for publication 27 Aug 2022. liage, such as baby leaf crops, must have et al., 2017). Pathovars are determined by host
Published online 17 Oct 2022. disease severity below 5% to satisfy producers
This research was made possible by funding
range and virulence (Baltrus et al., 2017). PSA
because of the difficulty of sorting symptomatic belongs to phylogroup 2 and is pathogenic on
support from the U.S. Department of Agricul-
ture Specialty Crop Research Initiative program
leaves (Pethybridge et al., 2018). Similarly cucurbits, such as watermelon as well as mem-
grant number 2019-51181-30019. Many thanks to stringent requirements may be found for Swiss bers of Amaranthaceae, such as table beet and
Dr. Carolee Bull and her graduate student Lindsay chard production. Additionally, table beets Swiss chard (Nikolic et al., 2018).
Boyd of The Pennsylvania State University and are often harvested by top-pulling machinery PSA can affect table beet and Swiss chard
Dr. Lindsey du Toit and her graduate student (Pethybridge et al., 2017, 2020). If greens are throughout the crops’ biennial lifecycle, in-
Marilen Nampijja of Washington State University too damaged by disease, the tops will be cluding seedlings, mature plants during vege-
for sharing their expertise and time to support pulled off the roots so that the roots will not tative growth, and flowering plants during
this work.
be harvested, leading to direct losses for growers reproductive growth (Jacobsen, 2009). How-
I.L.G. is the corresponding author. E-mail: ilgoldma@
wisc.edu. (Pethybridge et al., 2017, 2020). ever, in the vegetative phase, symptoms are
This is an open access article distributed under the In recent decades, a foliar disease known typically most severe early in growth or at up
CC BY-NC-ND license (https://creativecommons. as bacterial leaf spot (BLS) has increased in to eight sets of true leaves (Pethybridge et al.,
org/licenses/by-nc-nd/4.0/). severity across the United States, leading to 2018). During the second year of growth,

1436 HORTSCIENCE VOL. 57(11) NOVEMBER 2022

symptoms can occur throughout the season
from early on, when the foliage forms a compact
rosette, through the end of the season when
seeds are drying in the field. It is unknown
whether infection timing affects seed infec-
tion incidence, although seed infection po-
tentially has large negative impacts on the
ability to sell seed in addition to the cost of
treating seed. The BLS pathogen can be
seedborne, and infected seeds are one of
the main ways this pathogen is spread (Ark
and Leach, 1946; Gitaitis and Walcott, 2008;
Lamichhane et al., 2015). Infected seeds can
disseminate this pathogen across large geo-
graphic distances and are likely the main
method that BLS is spread to new locations
(Gitaitis and Walcott, 2008). The differences
in cultivation time between vegetable and
seed crops, combined with physiological dif-
ferences in the plants between the first and
second years of growth, have potentially
large implications for breeding programs
(Grahn et al., 2015).
Current BLS management strategies in-
clude preventative applications of copper treat-
ments, removal of crop residue, drip irrigation
to prevent splashing of the pathogen on leaves,
and crop rotation (Lamichhane et al., 2015;
Pethybridge et al., 2018). Crop rotation is a
limited strategy due to the number of host plant
species that PSA can both infect and survive
on epiphytically (Lamichhane et al., 2015;
Morris et al., 2013; Pethybridge et al., 2018;
Riffaud and Morris, 2002). Postharvest seed
treatment is another disease management op-
tion that can be effective for those seedborne
bacterial pathogens that colonize the seedcoat
rather than the endosperm, such as PSA (Singh
and Mathur, 2004). However, seed treatment is
expensive, and therefore, it is preferable to pro-
duce clean seed. In addition, table beet and
Swiss chard seed is often decorticated mechani-
cally before sale (Peck et al., 1967). Decortica-
tion has been shown to reduce the inoculum
load of some seedborne pathogens of beet and
chard without adverse effects on germination
(Peck et al., 1967; Singh and Mathur, 2004).
Depending on the environmental condi-
tions of a particular growing season, BLS
outbreaks can be severe and cause economic
losses. BLS thrives in wet and cool conditions
ranging from 10 to 25  C (Jacobsen, 2009; Pe-
thybridge et al., 2018). When conditions are
conducive, disease incidence and severity can
be high. For example, in New York in 2017,
when conditions were especially wet, the se-
verity of BLS in-table beet fields surveyed in
which the disease was observed was 75%,
whereas the average incidence of BLS in these
individual infected fields was 80% (S. Pethy-
bridge, Cornell University, personal communi-
cation). Similar patterns have been observed
in Washington State, with up to 50% inci-
dence and up to 75% severity observed in
some table beet seed crops in 2020, when con-
ditions were cool and wet. However, in 2015,
one of the hottest and driest years on record
for western Washington State, there was no
BLS observed in any of a dozen table beet
and Swiss chard seed crops surveyed (L. du
Toit, Washington State University, personal
HORTSCIENCE VOL. 57(11) NOVEMBER 2022
communication). Similar results were noted
during the hot and dry 2021 season in west-
ern Washington, when temperatures reached
40  C (L. du Toit, Washington State Univer-
sity, personal communication). In baby leaf
production, whole crops can be lost due to inci-
dences of disease as low as 5% because of the
difficulty of sorting symptomatic leaves after
harvest. For fresh market root or bunching
crops, some growers lose whole crops when
disease intensity is high.
As weather conditions change in beet and
chard growing areas, BLS impacts have the
potential to increase. In the midwestern and
northeastern United States, predicted in-
creases in precipitation, particularly in winter
and spring, may provide the cool, wet condi-
tions in the beginning of the growing season
under which P. syringae thrives. Such condi-
tions may also increase the chance of splash
dispersal within fields (Pryor et al., 2014; Xin
et al., 2018). The opposite rainfall patterns
are predicted in the Pacific Northwest and
southwestern United States (Gonzalez et al.,
2018; May et al., 2018). However, with drier
conditions expected in these areas, there will
be greater need for irrigation. Overhead irri-
gation increases the risk of splash dispersal of
this pathogen (Jacobsen, 2009). In baby leaf
production, planting densities are very high
(8 to 9 million seed/ha), and therefore,
splash dispersal can be detrimental. A study
by Derie et al. (2016) demonstrated that less
than 10 CFU/g seed is enough for seed trans-
mission of BLS under overhead irrigation in
baby leaf production in western Washington
State. Seed transmission at that low level of
seed infection was sufficient to render the
leaves unmarketable when conditions were
wet and cool (Derie et al., 2016).
Difficulties can arise when screening for
differences among accessions in a field set-
ting. For example, reports of BLS from field
surveys suggest there are usually “hot spots”
of disease resulting from natural infection,
meaning not all areas are affected equally
(L. du Toit, Washington State University, per-
sonal communication). A greenhouse setting
allows for better control over environmental
conditions, including temperature, humidity,
and soil moisture (Wintermantel and Kaffka,
2006). In addition, many studies in sugar
beets have demonstrated consistency between
greenhouse screens and field trials for ranking
of accessions between the two experimental
settings for several types of fungal diseases, in-
cluding black root rot and pocket rot patho-
gens, as well as viral diseases such as curly top
(Okazaki et al., 2005; Scholten et al., 2001;
Wintermantel and Kaffka, 2006).
Despite the potential impact BLS could
have on table beet and Swiss chard produc-
tion, at present, there are few highly effective
control strategies and no known genetic resis-
tance to this pathogen. On the basis of reports
from growers, field sampling, and known var-
iation among germplasm accessions to other
beet pathogens, we hypothesized that suscep-
tibility to BLS differs among B. vulgaris ac-
cessions. Therefore, the goal of this study
was to evaluate vegetable-type B. vulgaris
accessions, designated as plant introductions
(PIs), from the USDA National Plant Germ-
plasm System, publicly available inbred lines
from the University of Wisconsin–Madison
table beet breeding program, and commercial
cultivars of both table beet and Swiss chard
for their responses to PSA in a controlled
greenhouse setting to measure responses to
inoculation in both the vegetative and repro-
ductive growth stages.
Methods
Vegetative evaluations. In the spring and
summer of 2021, three experiments were
conducted to measure disease severity of dif-
ferent Beta vulgaris accessions when exposed
to PSA during vegetative growth. These are
hereafter called experimental repeats. A total
of 13 commonly grown table beet cultivars,
five widely used table beet breeding lines
from the University of Wisconsin–Madison
table beet breeding program, seven commonly
grown Swiss chard cultivars, and 27 PIs were
evaluated (Table 1).
PIs were evaluated in a group that is re-
ferred to hereafter as the PI screen. Commer-
cial cultivars and breeding lines were split
into two groups that are called the commer-
cial screens. Although all cultivars and breed-
ing lines were originally screened together,
lesions had a different physical appearance
on leaves with lighter colored foliage com-
pared with leaves with dark colored foliage.
The appearance of symptoms on cultivars
and breeding lines with light-colored foliage
was not always consistent with the descrip-
tion of lesions having tan to brown centers
surrounded by a dark border. In some acces-
sions, such as ‘Silverado’, lesions started out
tan to gray, and the dark border was absent
until later in the progression of the disease
(Supplemental Fig. 9). Therefore, accessions
with lighter colored foliage were re-screened.
Rescreening allowed for standardization of
disease ratings across all accessions. The re-
screening of the lighter colored accessions is
referred to as the commercial screen “light-
colored foliage accessions”; the screening of
the darker colored accessions is referred to as
the commercial screen “dark-colored foliage
accessions.” The duration of these vegetative
screens was 9 weeks from planting to final
evaluation.
Accessions within each group were orga-
nized in a completely randomized design in a
greenhouse at the Walnut Street Greenhouse
complex in Madison, WI. Greenhouse tables
were divided into 46 cm × 46 cm grids, with
each grid containing one pot. Approximately
five seeds were planted in a size 100 (11.75 cm
height, 11 cm top diameter; Nursery Supplies
Inc., Pairless Hills, PA) plastic pot in a one-to-
one mix of field soil from Madison, WI, and
ProMix (high porosity 1 bio fungicide and my-
corrhizae, Premier Tech Horticulture, Quebec,
Canada). Once plants exhibited one true leaf,
they were thinned to one plant per pot. Pots
were placed in saucers, and plants were bottom
watered via saucers to prevent splashing on the
leaves. Day length was set at 16 h with lighting
1437
Table 1. Accessions and sources for study of reaction of Beta vulgaris materials in response to inoculation with P. syringae.
Accession Source Accession type
Boro Johnny’s Selected Seeds Commercial Beet Cultivar
Bull’s Blood Johnny’s Selected Seeds Commercial Beet Cultivar
Chioggia Guardsmark Johnny’s Selected Seeds Commercial Beet Cultivar
Detroit Dark Red Reimer Seeds Commercial Beet Cultivar
Early Wonder Tall Top Johnny’s Selected Seeds Commercial Beet Cultivar
Golden Detroit Reimer Seeds Commercial Beet Cultivar
Merlin Territorial Seeds Commercial Beet Cultivar
Red Ace Reimer Seeds Commercial Beet Cultivar
Red Cloud Jung Seeds Commercial Beet Cultivar
Ruby Queen Reimer Seeds Commercial Beet Cultivar
Touchstone Gold Johnny’s Selected Seeds Commercial Beet Cultivar
Evansville Orbit UW Madison Carrot and Table Beet Laboratory Precommercial Beet Cultivar
Blushing Not Bashful UW Madison Carrot and Table Beet Laboratory Precommercial Beet Cultivar
W357B UW Madison Carrot and Table Beet Laboratory Breeding Line
W411A UW Madison Carrot and Table Beet Laboratory Breeding Line
W451C UW Madison Carrot and Table Beet Laboratory Breeding Line
W452C UW Madison Carrot and Table Beet Laboratory Breeding Line
W453B UW Madison Carrot and Table Beet Laboratory Breeding Line
Barese Johnny’s Selected Seeds Commercial Chard Cultivar
Bright Lights Johnny’s Selected Seeds Commercial Chard Cultivar
Fordhook Giant Johnny’s Selected Seeds Commercial Chard Cultivar
Magenta Sunset Johnny’s Selected Seeds Commercial Chard Cultivar
Rainbow Reimer Seeds Commercial Chard Cultivar
Rhubarb Johnny’s Selected Seeds Commercial Chard Cultivar
Silverado West Coast Seeds Commercial Chard Cultivar
Ames 22163 USDA National Plant Germplasm System, Pullman, WA PI
NSL 28020 USDA National Plant Germplasm System, Pullman, WA PI
NSL 28024 USDA National Plant Germplasm System, Pullman, WA PI
NSL 28026 USDA National Plant Germplasm System, Pullman, WA PI
PI 109039 USDA National Plant Germplasm System, Pullman, WA PI
PI 124528 USDA National Plant Germplasm System, Pullman, WA PI
PI 141919 USDA National Plant Germplasm System, Pullman, WA PI
PI 164292 USDA National Plant Germplasm System, Pullman, WA PI
PI 164805 USDA National Plant Germplasm System, Pullman, WA PI
PI 169015 USDA National Plant Germplasm System, Pullman, WA PI
PI 169028 USDA National Plant Germplasm System, Pullman, WA PI
PI 174059 USDA National Plant Germplasm System, Pullman, WA PI
PI 193458 USDA National Plant Germplasm System, Pullman, WA PI
PI 222234 USDA National Plant Germplasm System, Pullman, WA PI
PI 269309 USDA National Plant Germplasm System, Pullman, WA PI
PI 271439 USDA National Plant Germplasm System, Pullman, WA PI
PI 285591 USDA National Plant Germplasm System, Pullman, WA PI
PI 323938 USDA National Plant Germplasm System, Pullman, WA PI
PI 357357 USDA National Plant Germplasm System, Pullman, WA PI
PI 379097 USDA National Plant Germplasm System, Pullman, WA PI
PI 592989 USDA National Plant Germplasm System, Pullman, WA PI
PI 612330 USDA National Plant Germplasm System, Pullman, WA PI
PI 612334 USDA National Plant Germplasm System, Pullman, WA PI
PI 612338 USDA National Plant Germplasm System, Pullman, WA PI
PI 612340 USDA National Plant Germplasm System, Pullman, WA PI
W6 22192 USDA National Plant Germplasm System, Pullman, WA PI
PI 5 plant introduction; USDA 5 U.S. Department of Agriculture; UW 5 University of Washington.

in the greenhouse, and temperatures ranged
from 22 to 24  C. For each accession, four
plants were spray-inoculated with PSA and one
plant served as a negative control treatment that
was sprayed with sterilized Milli-Q water.
Plants were inoculated with PSA strain BP
1452 obtained from Dr. Carolee Bull of Penn-
sylvania State University that was isolated origi-
nally from Swiss chard in Washington State by
Dr. Lindsey du Toit. BP1452 is pathogenic on
both table beet and Swiss chard. The strain was
grown on nutrient agar in petri dishes and incu-
bated at 28  C with no light for 48 h. The spray
inoculation protocol was based on a modified
protocol from Dr. Carolee Bull. Briefly, colonies
of BP 1452 were suspended in sterilized MilliQ
water at a concentration of 1 × 108 CFU/mL
(optical density of 0.8 at 600 nm measured spec-
trophotometrically) directly before inoculation.
Inoculations occurred at the two true-leaf growth
stage and then again 2 weeks later. At each inoc-
ulation, each plant was sprayed with 4.7 mL of
inoculum or MilliQ water. The spray was tar-
geted at the underside of leaves on all sides of
the plant. Plants were enclosed in clear plastic
bags 24 h before inoculation and then re-
bagged for 48 h post-inoculation to increase
humidity and favor infection.
Visual disease ratings were conducted four
times per plant starting 1 week post-inocula-
tion. Ratings included the percentage surface
area of a pair of leaves with lesions (Table 2).
The first pair of leaves was the oldest pair.
This percentage was then translated into a leaf
pair score that ranged from 0 to 7 (Fig. 1).
This leaf pair score was then combined with
the number of leaf sets that had symptoms for
an overall plant score (Table 1). For example,
multiple sets of leaves for one plant individually
could have a score of 1, whereas the overall
plant has a score of 2 because the symptoms
were on multiple leaf pairs. Overall plant score
ranged from 0 to 7. For example, a score of
3 means that lesions started to coalesce, and a
score of 5 means that at least half of the first
two pairs of leaves died, and a leaf pair had at
least scores of 2 or higher.
Reproductive evaluations. Twenty-four ac-
cessions, including seven Swiss chard com-
mercial cultivars, 12 table beet commercial
cultivars, and five table beet breeding lines,
were sown by hand in the field during Sum-
mer 2020 at East Madison Agricultural Re-
search Station in Hartland, WI. Each accession
was represented by a 3.7-m-long row and was
replicated four times in the field. The layout
was a completely randomized design. Roots

1438 HORTSCIENCE VOL. 57(11) NOVEMBER 2022

Table 2. Bacterial leaf spot severity rating system of Beta vulgaris subsp. cicla and Beta vulgaris and threshed to prepare them for seed testing.
subsp. vulgaris inoculated with the bacterium Pseudomonas syringae pathovar aptata. Seeds from each plant were sent to Eurofins
Leaf set or Leaf set or Overall BioDiagnostics Inc. (Longmont, CO) and
individual individual leaf Description of overall plant plant tested for the presence of PSA. Seeds were
leaf percentage percentage score score-vegetative scorei sown in individual boxes. Any lesions that ap-
0% 0 Whole plant is healthy; no lesions 0 peared on the seedlings were then excised,
1% to 10% 1 Onset of disease: appearance of lesions on oldest 1 macerated, and plated onto semiselective
leaf set (leaf set 1) KBBC and KBZ media. Colonies were in-
10% to 25% 2 Increase in number of lesions on oldest leaves; 2 cubated for 4 to 7 d at 27 to 30  C. The col-
oldest sets of leaves (leaf sets 1 and 2) show onies were then compared with a positive
typical symptoms
control strain of PSA based on appearance.
25% to 50% 3 Spots merging (individual lesions no longer clearly 3
visible); large necrotic areas forming on oldest set No colonies matched the positive control.
50% to 75% 4 Lesions progressed onto newer leaves (sets 3 and 4 Therefore, no isolates were sequenced for
4); oldest set dead confirmation.
75% to 90% 5 At least half or more of the oldest leaves dead 5 Statistical analysis. Data from both the
(first and second set of leaves); more lesions on vegetative and reproductive evaluations were
newer leaves (third and fourth set) analyzed using R version 4.1.1 (RStudio, Bos-
90% to 99% 6 Oldest leaves (first and second set) dead; large 6 ton, MA). Analysis of variance (ANOVA)
necrotic areas on newer leaves (third and fourth
set)
models for vegetative and reproductive evalua-
100% (leaf dead) 7 First, second, and third set of leaves dead; fourth 7 tions were linear mixed models with accession
set large necrotic regions; fifth set has lesions and days post-inoculation as main effects, and
i
Overall plant score is based on the leaf set percentage score as well as the number of leaf sets with pot number as the random effect. These linear
symptoms for evaluations of plants in the vegetative stage of growth. mixed models were fit with ‘lmer’ from the
“lme4” package (version 1.1–23). ANOVA
was performed using the “lmerTest” package
were harvested 12 weeks after planting. At were enclosed in clear plastic bags 24 h before in R (version 3.1–2). When an interaction was
harvest, foliage was removed from the roots, inoculation and bagged again for 48 h post-in- significant between accession and days post-in-
leaving 2 cm of meristematic tissue at the oculation to increase humidity. Holes were cut oculation, results for each week were analyzed
crown. Roots were then packed with wood- in the tops of the bags through which the inocu- separately. In such cases, the model was a lin-
chips inside paper bags, which were then lum was sprayed onto all surfaces of the plant, ear fixed model with the main effect of acces-
placed inside plastic bags. The bags were then and then these holes were later sealed. sion, and data were fit using ‘lm’ from the
placed in a cooler at 12  C for vernalization. Disease ratings were conducted starting “stats” package (version 4.0.4).
After 13 weeks, roots were removed from the 1 week post-inoculation and occurred for ANOVAs were conducted with overall
cooler and planted in ProMix in size 200 pots the next 5 weeks. Ratings were based on the mean disease score from the four inoculated
(14.8 cm height, 14 cm top diameter) (Nursery percentage of leaf area with lesions, as de- plants to assess differences among accessions
Supplies Inc., Pairless Hills, PA) at the Horti- scribed earlier, with the exception that the in reaction to inoculation. ANOVAs were per-
culture Research Farm greenhouses (Arlington, percentage leaf area with symptoms was de- formed without data for the noninoculated con-
WI) in December. Pots were placed in saucers termined for individual leaves rather than leaf trol treatments because little disease was
and plants were watered from the saucers. Day pairs as leaf pairs were difficult to distinguish observed on these plants. Based on the
length was set at 16 h, and temperatures ranged at this growth stage. Four individual leaves ANOVA, all models with P values < 0.1
from 22 to 24  C. Flower stalks began to were chosen at random at the first rating and were analyzed with “emmeans” from the
emerge in January and were in full flower by marked to ensure ratings of the same leaf “emmeans” package (version 1.4-8) for de-
early February. week after week. Leaf scores for all four termining differences among accessions.
Two reproductive growth stage screens leaves were then averaged to give the overall Area under the disease progress curve
were conducted, and the duration of each plant score, which ranged from 0 to 7. A score (AUDPC) for each accession was estimated
screen was 12 weeks from planting of the of 0 meant that the plant was healthy (no le- using the “agricolae” package (version 1.3-3)
root until the final evaluation. These plants sions present). A score of 2 meant that the in- in R. AUDPC was used to compare acces-
were also inoculated with PSA strain BP dividual scores of the four leaves averaged sions as a quantitative measure of disease in-
1452. Preparation of bacteria and greenhouse together were a 2 or that, on average, the four cidence over the duration of the trial. AUDPC
layout were as described for the vegetative leaves had 10% to 25% of their total surface was analyzed as a fixed model with accession
experiments. Approximately 6 weeks after area with lesions. A score of 7 translated to all as the main effect. ANOVA of AUDPC values
planting roots, and then biweekly for the next four leaves being covered with lesions or was performed with “aov” from the “stats”
month, plants were spray-inoculated. Inoculum dead. package in R (version 4.0.4). Noninoculated
was prepared as described for the vegetative After 6 weeks of ratings, plants that were control treatments were excluded from the
screens. Each plant received 10.9 mL of the producing seed continued to be watered for 8 ANOVA analysis. Pairwise comparisons were
BP1452 suspension or MilliQ water. Plants more weeks. At this time, seeds were harvested made using “emmeans” at a 5 0.05.

Fig. 1. Disease rating scale for the percentage of vegetative leaf set (pair) and the reproductive growth stage leaf with symptoms of bacterial leaf spot for
Beta vulgaris subsp. cicla and Beta vulgaris subsp. vulgaris spray inoculated with Pseudomonas syringae pathovar aptata strain BP 1452. 0 5 0% dis-
eased tissue; 1 5 1% to 10% of leaf area with disease symptoms; 2 5 10% to 25% of leaf area affected; 3 5 25% to 50% of leaf area affected; 4 5 50%
to 75% of leaf area affected; 5 5 75% to 90% of leaf area affected; 6 5 90% to 99% of leaf area affected; 7 5 100% of leaf area affected/leaf is dead.

HORTSCIENCE VOL. 57(11) NOVEMBER 2022 1439


Fig. 2. Mean area under the disease progress curve (AUDPC) of eight Beta vulgaris subsp. vulgaris
commercial cultivars, one Beta vulgaris subsp. vulgaris breeding line, and one Beta vulgaris subsp.
cicla commercial cultivar in response to spray inoculation with Pseudomonas syringae pathovar
aptata strain BP 1452 during vegetative growth in two screening trials in a greenhouse in 2021.
Results
The method of inoculation was verified
by the presence of negative, noninoculated
control plants of each accession. Although
BLS was observed on some of the noninocu-
lated plants, there were large differences in
overall mean disease scores observed between
inoculated and noninoculated control plants for
all accessions (Supplemental Figs. 1–4), pro-
viding assurance that the inoculation technique
was suitable for comparison of accessions.
Vegetative evaluations, commercial screen:
dark-colored foliage accessions. No significant
interaction was found between experimental
repeat and accession for disease severity rat-
ings (P > 0.1). Therefore, data were analyzed
together for both experimental repeats. Addi-
tionally, no significant interaction was found
between days post-inoculation and accession
for BLS severity ratings. Significant differ-
ences were not detected among accessions
for overall disease ratings. Nevertheless,
‘Bull’s Blood’ consistently had one of the
lowest overall disease scores over time,
whereas ‘Boro’ consistently had one of the
highest over time. BLS symptoms occurred
on all inoculated plants in experimental repeats
1 and 2. The mean AUDPC ratings for inocu-
lated plants ranged from 3.94 to 7.36 with an
average AUDPC of 5.70 (Fig. 2). No symp-
toms occurred on six of 10 control plants in ex-
perimental repeat 1, and the mean AUDPC
was 0.70. ANOVA revealed no effect of acces-
sion on mean AUDPC (P > 0.1), although
‘Bull’s Blood’ had the smallest AUDPC, and
‘Boro’ and ‘Detroit Dark Red’ had the largest
AUDPCs (Fig. 2).
Commercial screen: light-colored foliage
accessions. BLS symptoms occurred on all
inoculated plants in experimental repeats 1
and 2. For experimental repeat 1, the mean
AUDPC rating for inoculated plants ranged
from 1.62 to 8.5 with an average AUDPC of
4.92 (Figs. 3 and 4). For experimental repeat
two, the mean AUDPC for inoculated plants
ranged from 4.5 to 11.67 with an average
AUDPC of 8.77 (Figs. 3 and 4). No symptoms
occurred on 11 of 16 control plants in experi-
mental repeat 1. The mean AUDPC was 0.66.
In repeat 2, eight of 16 control plants did not
have symptoms, and the mean AUDPC was
1.03.
ANOVA of disease severity ratings for the
combined experimental repeat revealed a sig-
nificant interaction for experimental run and
accession (P < 0.01). Therefore, experimental
repeats were kept separate for analysis. ANOVA
for experimental repeat 1 showed a significant
interaction between accession and days post-
inoculation on overall mean disease score
(P 5 0.05), indicating that accessions did not
maintain the same ranking at each rating time
(Table 3). At 7 and 14 d post-inoculation, acces-
sion had no significant effect on overall
mean disease score (P > 0.05) because there
was inadequate disease intensity to differenti-
ate among accessions (Table 3; Fig. 3). AN-
OVA of overall mean disease scores 21 d post-

Fig. 3. Area under the disease progress curve (AUDPC) of six Beta vulgaris subsp. vulgaris commercial cultivars, four Beta vulgaris subsp. vulgaris breed-
ing lines, and six Beta vulgaris subsp. cicla commercial cultivars in response to spray inoculation with Pseudomonas syringae pathovar aptata strain BP
1452 during vegetative growth in two greenhouse screening trials in 2021.

1440 HORTSCIENCE VOL. 57(11) NOVEMBER 2022


Fig. 4. Mean area under the disease progress curve (AUDPC) of six Beta vulgaris subsp. vulgaris com-
mercial cultivars, four Beta vulgaris subsp. vulgaris breeding lines, and six Beta vulgaris subsp. ci-
cla commercial cultivars in response to spray inoculation with Pseudomonas syringae pathovar
aptata strain BP 1452 during vegetative growth in a greenhouse screen carried out twice in 2021.
Bars not sharing a common letter are significantly different at P # 0.05.
inoculation indicated significant differences
among accessions (P < 0.05) (Table 2).
‘Blushing Not Bashful’ and ‘Touchstone Gold’
had significantly lower mean disease scores
than ‘Golden Detroit’, ‘Silverado’, ‘Chioggia
Guardsmark’, ‘Detroit Dark Red’, ‘Barese’,
‘Bright Lights’, ‘Rhubarb’, and W451C. The
main effect of accession also was significant
for overall mean disease score (P < 0.05) 28 d
post-inoculation (Table 3). W452C and ‘Blushing
Not Bashful’ had significantly lower mean disease
scores than ‘Silverado’ and ‘Chioggia Guards-
mark’. ‘Touchstone Gold’ had a significantly lower
mean disease score than ‘Silverado’, ‘Chioggia
Guardsmark’, ‘Golden Detroit’, W411A, ‘Bright
Lights’, ‘Detroit Dark Red’, ‘Rhubarb’, W451C,
‘Barese’, and ‘Evansville Orbit’.
ANOVA for experimental repeat 2 also
showed a significant interaction between ac-
cession and days post-inoculation on overall
mean disease score (P < 0.01). Therefore,
the effect of accession was analyzed for each
week individually. At 7 d post-inoculation,
HORTSCIENCE VOL. 57(11) NOVEMBER 2022
accession had a significant effect on mean
overall mean disease score (P < 0.05) be-
cause BLS developed much quicker in repeat
2 than in repeat 1 (Table 3; Fig. 3). Acces-
sions ‘Rainbow’, ‘Touchstone Gold’,
‘Fordhook Giant’, ‘Chioggia Guardsmark’,
and ‘Golden Detroit’ all had significantly
lower mean overall disease ratings compared
with ‘Silverado’ and W451C at this 7-day
rating (Table 3). Accession also had a signifi-
cant effect on the overall mean disease score 14
d post-inoculation (P < 0.05) (Table 3), when
‘Chioggia Guardsmark’ and ‘Golden De-
troit’ had significantly lower mean overall
disease scores than the other accessions. AN-
OVA of overall mean disease scores 21 or 28 d
post-inoculation indicated no significant differ-
ence among accessions (Table 3; Supplemental
Figs. 5–8).
ANOVA for experimental repeat one showed
a significant effect of accession on mean
AUDPC (P < 0.05). ‘Touchstone Gold’ had
a significantly smaller AUDPC compared with
‘Silverado’, ‘Rhubarb’, ‘Chioggia Guardsmark’,
‘Bright Lights’, W451C, ‘Detroit Dark Red’, and
‘Golden Detroit’ (Fig. 4). AUDPC for experi-
mental repeat 2 showed a significant effect of
accession on mean AUDPC (P < 0.1). The
mean AUDPC value for ‘Golden Detroit’
and ‘Chioggia Guardsmark’ was signifi-
cantly less than the mean AUDPC for
‘Evansville Orbit’, W453B, W411A, W451C,
‘Rhubarb’, or ‘Silverado’, in contrast to results
for repeat 1 of this experiment (Fig. 4, Supple-
mental Figs. 5–8). In both repeat 1 and repeat
2, ‘Touchstone Gold’ consistently had one of
the smallest AUDPCs. W451C consistently
had one of the largest AUDPCs in both repeats.
‘Rainbow’ was consistently the Swiss chard
cultivar with the smallest AUDPC rating. No
Swiss chard cultivar reliably had the largest
AUDPC rating of the accessions evaluated in
these trials.
PI screen. BLS symptoms occurred on all
inoculated plants in experimental repeats 1
and 2. For experimental repeat 1, the mean
AUDPC rating for inoculated plants ranged
from 1.33 to 8.75 with an average AUDPC
rating of 5.35 (Figs. 5 and 6). For experimen-
tal repeat 2, the mean AUDPC for inoculated
plants ranged from 1.5 to 8.75 with an aver-
age AUDPC rating of 4.37 (Figs. 5 and 6).
No symptoms occurred on eight of 25 control
plants in experimental repeat 1, and the mean
AUDPC rating was 2.26. In repeat 2, 15 of
20 control plants did not have symptoms, and
the average AUDPC rating was 0.78.
Significant interactions were found for ex-
perimental repeat and accession on BLS disease
severity ratings (P < 0.001). In experimental
repeat 1, plants exhibited higher overall mean
disease scores than plants in experimental re-
peat 2. Therefore, experimental repeats were
kept separate for analysis. There was no sig-
nificant interaction between accession and
days post-inoculation for either experimental
repeat (Table 4).
ANOVA for 7 d post-inoculation indicated
that accession had no significant effect on over-
all mean disease score (P > 0.1) (Table 4;
Supplemental Fig. 7). ANOVA at 14 d post-in-
oculation showed a moderately significant dif-
ference between in overall mean disease scores
among the accessions (P < 0.1) (Table 4). PI
222234 and PI 174059 had a significantly
smaller overall mean disease score compared
with PI 164805, PI 612338, NSL 28020, PI
285591, PI 169015, PI 124528, and PI 379097.
ANOVA at 21 d (P < 0.05) and 28 d (P < 0.1)
post-inoculation were also significant for the
effect of accession on mean disease ratings. At
21 d post-inoculation, PI 222234 and PI 174059
had significantly smaller overall mean disease
scores than Ames 22163, PI 323938, NSL
28024, PI 169028, PI 612330, PI 164805, PI
285591, PI 141919, PI 169015, NSL 28020, PI
612338, PI 379097, and PI 124528. Finally, at 28
d post-inoculation, PI 222234, PI 174059, and PI
271439 had significantly smaller overall mean
disease scores than PI 169015, PI 141919, NSL
28020, PI 379097, PI 612338, and PI 124528.
ANOVA for experimental repeat 2 at 7
and 14 d post-inoculation did not show a sig-
nificant difference in mean disease scores due
1441
Table 3. Analysis of variance for experimental runs 1 and 2 for average disease scores of six Beta been completed before this recognition, thus
vulgaris subsp. cicla commercial cultivars, six Beta vulgaris subsp. vulgaris commercial cultivars, necessitating dropping the light-colored fo-
and four Beta vulgaris subsp. vulgaris breeding lines in response to inoculation with Pseudomonas liage accessions from the analysis.
syrinage pathovar aptata strain BP 1452 in a greenhouse screen evaluated in 2021.
ANOVA of experimental repeats 1 and 2
Run 1: source of variation Numerator df Denominator df Mean squares Significance combined did not reveal a significant inter-
Accession 15 40 0.83 ** action between experimental repeat and ac-
7 d post-inoculation 0.88 NS cession. Experimental repeat was moderately
14 d post-inoculation 1.2 NS significant, however (P < 0.1), with repeat 2
21 d post-inoculation 1.64 ** generally having a higher overall mean disease
28 d post-inoculation 1.93 ** score than repeat 1. As a result, repeat trial
Days post-inoculation 3 120 33.15 **** data were analyzed separately.
Accession: days post-inoculation 45 120 0.54 **
ANOVA for experimental repeat 1 did not
Run 2: Source Numerator df Denominator df Mean squares Significance reveal a significant interaction between acces-
Accession 15 39.10 0.39 ** sion and days post-inoculation (Table 5). No
7 d post-inoculation 1.32 ** rating days had significant differences in over-
14 d post-inoculation 1.23 ** all mean disease scores as a result of accessions
21 d post-inoculation 0.64 NS
(Table 5). ANOVA for experimental repeat 2
28 d post-inoculation 0.69 NS
Days post-inoculation 3 115.31 24.85 **** indicated a significant interaction between ac-
Accession: days post-inoculation 45 115.31 0.39 *** cession and days post-inoculation (P < 0.05)
NS, *, **, ***, **** Nonsignificant or significant at P # 0.1, 0.05, 0.01, or 0.001, respectively. (Table 5). Accessions changed rank as days
post-inoculation increased. ANOVA indicated
no significant differences in overall mean dis-
ease as a result of accessions, regardless of rat-
to accessions (Table 4). At 21 d post-inocula- PI 141919, PI 285591, PI 612338, NSL 28020, ing period (Table 5; Supplemental Fig. 8).
tion, accessions had a significant effect on PI 169015, PI 124528, and PI 379097 (Fig. 6). BLS symptoms occurred on all inoculated
overall mean disease (P < 0.05) (Table 4). PI AUDPC rating for experimental repeat 2 plants in experimental repeats 1 and 2. For
193458, PI 222234, PI 109039, PI 169015, PI showed a moderately significant effect of experimental repeat 1, the mean AUDPC rat-
269309, PI 592989, PI 612334, and PI 323938 accession on mean AUDPC (P < 0.01). PI ing for inoculated plants ranged from 3.38 to
all had significantly smaller overall mean dis- 193458 had a significantly smaller AUDPC 4.84 with an average AUDPC rating of 4.23
ease scores than PI 169028, PI 271439, and PI rating than PI 164805, PI 271439, or NSL (Fig. 7). For experimental repeat 2, the mean
28020. At 21 d post-inoculation, accession 28020 (Fig. 6). PI 22234 and NSL 28026 AUDPC for inoculated plants ranged from
again was significant for overall mean disease performed consistently with some of the 3.19 to 6.44 with an average AUDPC rating
(P < 0.05) (Table 4). PI 109039, PI 269309, smallest AUDPCs in both experimental re- of 4.84 (Fig. 7). Overall, both repeats 1 and 2
PI 592989, PI 323938, PI 612334, PI 193458, peats. NSL 28020 had one the largest of the reproductive evaluations had a smaller
PI 222234, PI 169015, PI 141919, and PI AUDPC ratings in both repeats. PI 612330 range for AUDPC ratings than the vegetative
612340 all had significantly smaller overall and PI 612338 were consistently midrange evaluations. In experimental repeat 1, all non-
mean disease scores than PI 28020, PI for AUDPC ratings. inoculated control plants showed symptoms
169028, and PI 271439. Reproductive evaluations. Although re- of disease. The mean AUDPC rating was
ANOVA for experimental repeat 1 showed productive evaluations were conducted with 1.58. In repeat 2, one of nine control plants
a significant effect of accession on mean 24 accessions, only 10 accessions were in- did not have symptoms, and the average
AUDPC ratings (P 5 0.05). PI 222234 and PI cluded in the analysis because of differences AUDPC rating was 2.1.
174059 had significantly smaller AUDPC rat- between accessions with light- and dark-col- No significant effect of accession on
ings than PI 612330, PI 164805, NSL 28024, ored foliage. Reproductive evaluations had mean AUDPC rating was noted for experimental

Fig. 5. Area under the disease progress curve (AUDPC) of 26 Beta vulgaris subsp. vulgaris plant introductions (PIs) in response to spray inoculation with
Pseudomonas syringae pathovar aptata strain BP 1452 during vegetative growth in two greenhouse screening carried out twice in 2021.

1442 HORTSCIENCE VOL. 57(11) NOVEMBER 2022


Fig. 6. Mean AUDPC of 26 Beta vulgaris subsp. vulgaris plant introductions (PIs) in response to spray
inoculation with Pseudomonas syringae pathovar aptata strain BP 1452 during vegetative growth in
a greenhouse trial carried out twice in 2021. Bars not sharing a common letter are significantly dif-
ferent at P # 0.05.
repeat 1 or 2 (P > 0.1). Although there were
no significant differences among accessions,
some patterns emerged. ‘Kestrel’ had the
smallest AUDPC rating in both repeats. ‘Boro’
and W357B consistently were midrange for
AUDPC. Similar to the vegetative screen, ‘Red
Cloud’ consistently had one of the largest
AUDPC ratings.
Seed testing for the presence of PSA did
not result in positive identification of any in-
oculated plants despite these plants showing
leaf symptoms. Overall, seed set was low
compared with a typical seed production field
because plants were in individual pots and
widely spaced in the greenhouse. In addition,
moisture and humidity may not have been
optimal for the pathogen to colonize develop-
ing fruits and seeds.
Discussion
The method of inoculation and testing used in
this study revealed significant differences in leaf
ratings between inoculated and noninoculated
HORTSCIENCE VOL. 57(11) NOVEMBER 2022
control plants, suggesting this approach was valu-
able for evaluation of BLS in Beta vulgaris. The
noninoculated control plants served as a check to
indicate that symptoms were the result of inocula-
tion with PSA (Supplemental Figs. 1–4). How-
ever, it was unknown if seed used for the
vegetative and reproductive screens were clean of
PSA. The negative control plants of certain acces-
sions developed BLS lesions while others did not
display symptoms. This suggests that seeds for
these particular accessions may have been in-
fected with PSA or that there may have been a
low level of cross-contamination between plants
over the duration of each experiment. Future ex-
periments could either use verified clean seed or
treat the seed before use in disease screens to re-
duce this potential effect. These disease evalua-
tions also demonstrated that Beta vulgaris subsp.
vulgaris commercial cultivars, breeding lines, and
PIs as well as Beta vulgaris subsp. cicla commer-
cial cultivars have a range of reactions to spray
inoculation with PSA.
Disease screens during the vegetative
growth stage of beet or chard accessions with
light-colored foliage revealed significant differ-
ences among accessions in response to inocula-
tion with PSA. The main effect of accession in
experimental repeat 2 was marginally signifi-
cant with a P value of 0.065; however, some
individual comparisons were significant at
P < 0.01. For table beets, ‘Touchstone Gold’
showed the greatest resistance based on a con-
sistently low AUDPC in both repeats, whereas
inbred line W451C consistently had one of the
highest AUDPCs. In experimental repeat 1,
‘Touchstone Gold’ had a 78% smaller mean
AUDPC rating than ‘Detroit Dark Red’ and a
76% smaller mean AUDPC rating than
W451C. In experimental repeat 2, the differ-
ences were greatly reduced. ‘Touchstone
Gold’ had a 12% smaller mean AUDPC rating
than ‘Detroit Dark Red’ and a 29% smaller
mean AUDPC rating than W451C.
For Swiss chard, ‘Rainbow’ consistently had
the lowest mean AUDPC rating. ‘Silverado’,
‘Bright Lights’, and ‘Rhubarb’ were variable be-
tween experimental repeats but exhibited some
of the highest mean AUDPC ratings. Koike
et al. (2003) first reported BLS infection on
commercially grown Swiss chard in California
from 1999 to 2003. However, they did not in-
dicate the cultivar(s) affected, and little work
has been done since to determine which culti-
vars are more susceptible or if susceptibility
varies among cultivars. This study demon-
strates that there is variability in responses
among Swiss chard cultivars.
Generally, plants in experimental repeat 2
had more severe BLS ratings compared with
plants in repeat 1 for accessions with light-
colored foliage. Experimental repeat 1 oc-
curred from the end of June to the beginning
of August, whereas repeat 2 occurred from
the end of July to the beginning of Septem-
ber. Although both occurred during the sum-
mer months, it is possible that differences in
environmental conditions between these time
periods accounted for some of the differ-
ences. In addition, the experimental repeats
took place in different greenhouses, which
could have had differences in temperature
and/or humidity. Finally, experimental repeat
1 of the light-colored foliage accessions was
the first round of screening in which we were
aware of the difference in BLS symptoms on
chard vs. beet cultivars and could therefore
adjust the foliar ratings for this observation.
‘Silverado’ has been known to display BLS
symptoms and has been used as a susceptible
cultivar in other BLS screens (L. du Toit,
Washington State University, personal commu-
nication). In experimental repeat one and two,
‘Silverado’ consistently exhibited symptoms
validating the methods used in these screens.
‘Detroit Dark Red’ was included as a check in
the commercial screens of accessions with both
dark- and light-colored foliage. In all repeats,
‘Detroit Dark Red’ had relatively consistent
performance, with AUDPC scores ranking this
cultivar with the most susceptible accessions.
No significant differences were observed
among accessions with dark-colored foliage
during either the vegetative or reproductive
screens. Experimental repeat 1 of the reproduc-
tive screens had a mean AUDPC rating range of
1443
Table 4. Analysis of variance for experimental runs 1 and 2 for average disease scores of 26 Beta vul- beet cultivars with red pigmented roots. Some
garis subsp. vulgaris plant introduction in response to inoculation with Pseudomonas syringae of the F1 hybrid table beet cultivars available
pathovar aptata strain BP 1452 during vegetative growth in a greenhouse in 2021.
on the market make use of male sterile lines
Run 1: source of variation Numerator df Denominator df Mean squares Significance developed at the University of Wisconsin–
Accession 26 65 0.20 *** Madison from the 1960s to the present. Because
7 d post-inoculation 0.98 NS table beet is a relatively minor specialty crop in
14 d post-inoculation 1.05 * the United States that does not receive large
21 d post-inoculation 1.25 ** amounts of breeding activity, the amount of ge-
28 d post-inoculation 1.20 * netic diversity among red-rooted cultivars may
Days post-inoculation 3 195 3.18 **** be limited.
Accession: days post-inoculation 78 195 0.12 NS
The open-pollenated cultivar Ruby Queen
Run 2: source of variation Numerator df Denominator df Mean squares Significance performed consistently between reproductive ex-
Accession 25 57 0.20 * perimental repeats and was in the middle of the
7 d post-inoculation 0.82 NS range of mean AUDPC ratings for both repro-
14 d post-inoculation 0.94 NS ductive and vegetative evaluations. This is in con-
21 d post-inoculation 0.88 ** trast to an unpublished field study from Cornell
28 d post-inoculation 0.98 ** University that found that ‘Ruby Queen’ showed
Days post-inoculation 3 171 4.00 ****
Accession: days post-inoculation 75 171 0.13 NS
“low” BLS symptoms (S. Pethybridge, personal
communication). Our greenhouse study also
NS, *, **, ***, **** Nonsignificant or significant at P # 0.1, 0.05, 0.01, or 0.001, respectively.
included ‘Red Cloud’, which was considered
to have “moderate” disease levels in the Cor-
nell study. The reproductive and vegetative ex-
3.38 to 4.84, and repeat 2 had a mean rating range Dark Red’ and a 45% smaller mean AUDPC perimental repeats were consistent, with ‘Ruby
of 3.19 to 6.44. The range of mean AUDPC rat- rating than ‘Red Cloud’. Queen’ showing fewer disease symptoms than
ings for the vegetative evaluation was 3.94 to The lack of significance among darker ‘Red Cloud’. In a study by Pethybridge et al.
7.36. Although there were no statistical differ- color accessions inoculated with PSA may (2017), ‘Ruby Queen’ was the least susceptible
ences among accessions, some patterns emerged have resulted from several factors. As a result fresh market cultivar to Cercospora leaf spot
that could be examined in future studies. The ac- of the small sample size, standard errors were (CLS), one of the most widespread and damag-
cession with one of the largest AUDPC rating relatively large. Increasing the sample size ing foliar diseases for table beet and Swiss
was ‘Red Cloud’. However, an unpublished study for each accession would increase the power chard. ‘Ruby Queen performance in both CLS
from Sarah Pethybridge (Cornell University, per- to identify differences among accessions. and BLS disease evaluations makes it a candi-
sonal communication) showed that ‘Red Cloud’ Additionally, disease screens require not date for further study and breeding efforts.
had “moderate” BLS symptoms compared with only a suitable host and a virulent pathogen In this greenhouse study, PIs showed sig-
the cultivar Pablo, which showed severe disease. but suitable environmental conditions. In these nificant differences in responses to spray inoc-
One of the more resistant accessions in the repro- screens, plants were enclosed in plastic bag for ulation with PSA, which might be expected
ductive evaluations was ‘Kestrel’, although statis- 24 h pre-inoculation and 48 h post-inoculation. from this more diverse collection of germ-
tical analysis did not separate out this cultivar However, development of the pathogen would plasm compared with the commercial cultivars
from the others screened. In experimental repeat have benefitted from increased humidity for screened. Generally, the AUDPC for PIs
1, ‘Kestrel’ exhibited a 27% smaller AUDPC longer periods to initiate and sustain infection spiked shortly following inoculation and then
than ‘Red Cloud’. In experimental repeat 2, and disease development. Subsequent experi- leveled off or showed a gradual increase
‘Kestrel’ exhibited a 50% smaller mean AUDPC ments could benefit from increasing humidity through the remaining disease evaluation peri-
rating than ‘Red Cloud’. One of the most resis- through the duration of disease rating. A third ods. Although there was variability in mean
tant accessions in the vegetative evaluations possibility for the lack of significant differences AUDPC ratings between repeats 1 and 2 of
was ‘Bull’s Blood’, which exhibited a 47% among accessions with dark colored foliage is the screening, PI 222234 and NSL 28026
smaller mean AUDPC rating than ‘Detroit the relatively limited genetic variability of table consistently ranked among the lowest mean
AUDPC ratings for the PI accessions. PI
612338 and NSL 28020 consistently ranked
Table 5. Analysis of variance for experimental run 1 and 2 for average disease scores of one Beta among the highest mean AUDPC ratings. PI
vulgaris subsp. cicla commercial cultivar, seven Beta vulgaris subsp. vulgaris commercial culti-
22234 had only 18% of the mean AUDPC
vars, and one Beta vulgaris subsp. vulgaris breeding lines in response to inoculation with Pseudo-
monas syringae pathovar aptata strain BP 1452 during reproductive growth in a greenhouse in value of NSL 28020 in experimental repeat
2021. 1 and 34% of the mean AUDPC value of
NSL 28020 in experimental repeat 2. NSL
Run 1: source df Mean squares Significance 28026 had 47% of the mean AUDPC value
Accession 9 0.01 NS of NSL 28020 in experimental repeat 1 and
7 d post-inoculation 0.16 NS 17% of the mean AUDPC value of NSL
14 d post-inoculation 0.10 NS 28020 in experimental repeat 2. PI 109039,
21 d post-inoculation 0.05 NS Ames 22163, PI 269309, and PI 592989
28 d post-inoculation 0.08 NS
35 d post-inoculation 0.1 NS
also had relatively stable performance be-
42 d post-inoculation 0.08 NS tween both repeats with mean AUDPC rat-
Days post-inoculation 5 3.33 **** ings on the smaller end of the range.
Accession: days post-inoculation 45 0.07 NS Testing of the seed harvested from the
Run 2: source df Mean squares Significance plants in the reproductive trials in this study,
using seedling grow outs by a commercial
Accession 8 0.04 NS
seed testing laboratory, did not identify any
7 d post-inoculation 0.14 NS
14 d post-inoculation 0.18 NS seed-borne PSA despite seed coming from
21 d post-inoculation 0.20 NS plants with BLS lesions. During seed produc-
28 d post-inoculation 0.22 NS tion in a field environment, the microclimate
35 d post-inoculation 0.34 NS can be conducive to BLS due to dense canopies
Days post-inoculation 5 2.92 **** that increase the humidity in the canopy, and
Accession: days post-inoculation 40 0.09 ** splash dispersal from rain or irrigation can eas-
NS, *, **, ***, **** Nonsignificant or significant at P # 0.1, 0.05, 0.01, or 0.001, respectively. ily spread the pathogen between neighboring

1444 HORTSCIENCE VOL. 57(11) NOVEMBER 2022

Fig. 7. Area under the disease progress curve (AUDPC) of eight Beta vulgaris subsp. vulgaris commercial cultivars, one Beta vulgaris subsp. vulgaris breed-
ing line, and one Beta vulgaris subsp. cicla commercial cultivar in response to spray inoculation with Pseudomonas syringae pathovar aptata strain BP
1452 during reproductive growth in a greenhouse trial completed twice in 2021.
plants. Therefore, experiments to look at seed
infection should occur in a field setting more
representative of production conditions. Addi-
tionally, future seed testing could entail a seed
wash technique that might be more effective at
detecting seedborne PSA than grow-out assays.
There was no clear correlation for BLS
severity between vegetative and reproductive
evaluations of beet and chard accessions in
this study. Depending on which experimental
repeats were compared, Pearson’s correlation
coefficients ranged from almost 0 to 0.36 for
all accessions and from 0.06 to 0.78 for table
beet cultivars. This suggests that the correla-
tion between vegetative and reproductive re-
sponses to spray inoculation was inconsistent.
The relationship between disease reaction in
vegetative and reproductive phases should be
addressed in future studies with a larger sam-
ple size for each accession. The significance
of this correlation is in the degree to which
plant breeders might need to screen at both the
vegetative and reproductive phases of growth
when breeding for resistance to BLS. If subse-
quent experiments demonstrate a significant
correlation between these two stages of the
plant life cycle, it may be possible to select
only in the vegetative phase, thereby saving
nearly 1 year per cycle during the breeding
process. If the disease responses of accessions
during two stages of the life cycle are not cor-
related, breeders may need to screen in both
stages.
PI accessions were expected to have
greater variability in response to spray inocu-
lation with PSA than commercial cultivars as
PIs commonly have a broad range of variabil-
ity for agronomic traits (Wigg and Goldman,
2020). The PI accessions did have the largest
range of mean AUDPC ratings in both re-
peats; however, the range in mean AUDPC
ratings for the commercial screen of light-
colored foliage accessions was similar. This
contrasts with findings by Wigg and Gold-
man (2020) for Rhizoctonia root rot of table
HORTSCIENCE VOL. 57(11) NOVEMBER 2022
beet, in which PIs had less variation com-
pared with commercial cultivars when inocu-
lated with Rhizoctonia solani. However,
when looking at the accessions with light-col-
ored foliage, inbreds had less variation than
commercial cultivars. The commercial screen
of accessions with light-colored foliage also
showed that table beets had a greater range of
mean AUDPC ratings than Swiss chard.
It should be noted that the screen of
light-colored foliage accessions of commer-
cial germplasm only included inbreds and
open-pollinated cultivars. The screen for dark-
colored foliage accessions of commercial germ-
plasm included both open-pollinated and hybrid
cultivars as well as one inbred. Hybrids had a
slightly smaller AUDPC range, 5.00 to 7.43
compared with open-pollinated cultivars, which
had a range of 3.94 to 7.93. The screening with
the smallest range for AUDPC ratings in both
experimental repeats were the reproductive
evaluations. The reproductive evaluations in-
cluded both open-pollinated and hybrid cultivars
as well one as one inbred. There was no clear
distinction between hybrid or open-pollinated
cultivars regarding range of AUDPC ratings.
Public knowledge about the relative sus-
ceptibility of B. vulgaris cultivars or acces-
sions to BLS has only started to emerge in
recent years. This study serves to build our
knowledge of how PSA effects B. vulgaris,
including B. vulgaris subsp. vulgaris com-
mercial cultivars, breeding lines, and PIs
and B. vulgaris subsp. cicla commercial cul-
tivars. Overall, this study identified a range
of B. vulgaris accessions and cultivar re-
sponses to inoculation with PSA, suggesting
varying susceptibility to BLS. On the basis
of overall mean disease scores and AUDPC
ratings, table beet and Swiss chard cultivars
recommended for future studies and breeding
are ‘Touchstone Gold’, ‘Bull’s Blood’, ‘Ruby
Queen’, ‘Kestrel’, and ‘Rainbow ‘because of
their lower BLS ratings. Several PIs, including
PI 222234 and NSL 28026, appear to have the
potential to offer valuable resistance to BLS to
be used in future breeding efforts. Further-
more, these, along with more susceptible lines,
can be used to generate mapping populations
to identify important regions controlling resis-
tance to PSA. Development of BLS resistant
table beets and Swiss chard will aid producers
in controlling this growing threat.
References
Arabiat, S., A.K. Chanda, K. Chittem, and M.F.R.
Khan. 2016. First report of Pseudomonas sy-
ringae pv. aptata causing bacterial blight of
sugar beet (Beta vulgaris) in Oregon. Plant
Dis. 100(11):2334, https://doi.org/10.1094/pdis-05-
16-0746-pdn.
Ark, P.A. and L.D. Leach. 1946. Seed transmission
of bacterial blight of sugar beet. Phytopathol-
ogy 36:549–553.
Baltrus, D.A., H.C. McCann, and D.S. Guttman. 2017.
Evolution, genomics, and epidemiology of Pseudo-
monas syringae. Mol. Plant Pathol. 18(1):152–168,
https://doi.org/10.1111/mpp.12506.
Brown, N.A. and C.O. Jamieson. 1913. A bacte-
rium causing a disease of sugar beet and nas-
turtium leaves. J. Agric. Res. 1:189–210.
Carsner, E. 1944. Black streak, a bacterial disease
of sugar beet in the Pacific Northwest. Phyto-
pathology 34:933–934.
Derie, M.L., B.J. Holmes, I. Safni, C.T. Bull, and
L.J. du Toit. 2016. Seedborne inoculum
thresholds of Pseudomonas syringae pv. ap-
tata, causal agent of bacterial leaf spot, in
‘Baby Leaf’ Swiss chard crops. Phytopathol-
ogy 106(Suppl 4):142.
Dutta, B., T. Ingram, R.D. Gitaitis, D.B. Langston,
T. Brenneman, T.M. Webster, and R.F. Davis.
2014. First report of bacterial blight of sugar
beet caused by Pseudomonas syringae pv. ap-
tata in Georgia, USA. Plant Dis. 98(10):1423,
https://doi.org/10.1094/PDIS-03-14-0235-PDN.
Gitaitis, R. and R. Walcott. 2008. The epidemiol-
ogy and management of seedborne bacterial
diseases. Annu. Rev. Phytopathol. 45:371–397,
https://doi.org/10.1146/annurev.phyto.45.06280
6.094321.
Gomila, M., A. Busquets, A. Mulet, E. Garcıa-
Valdes, and J. Lalucat. 2017. Clarification of
1445
 taxonomic status within the Pseudomonas sy-
ringae species group based on a phylogenomic
analysis. Front. Microbiol. 8:2422, https://doi.
org/10.3389/fmicb.2017.02422.
Gonzalez, P., G.M. Garfin, D.D. Breshears, K.M.
Brooks, H.E. Brown, E.H. Elias, A. Gunasekara,
N. Huntly, J.K. Maldonado, N.J. Mantua, H.G.
Margolis, S. McAfee, B.R. Middleton, and B.H.
Udall. 2018. Southwest, p. 1101–1184. In:
D.R. Reidmiller, C.W. Avery, D.R. Easterling,
K.E. Kunkel, K.L.M. Lewis, T.K. Maycock,
and B.C. Stewart (eds.). Impacts, risks, and ad-
aptation in the United States: Fourth national
climate assessment, Volume II. U.S. Global
Change Research Program, Washington, DC.
https://doi.org/10.7930/NCA4.2018.CH25.
Grahn, C.M., C. Benedict, T. Thornton, and C. Miles.
2015. Production of baby-leaf salad greens in the
spring and fall seasons of northwest Washington.
HortScience 50(10):1467–1471, https://doi.org/
10.21273/hortsci.50.10.1467.
Jacobsen, B.J. 2009. Bacterial leaf spot, p. 59–60. In:
R.M. Harveson, L.E. Hanson, and G.L. Hein (eds.).
Compendium of beet diseases and pests. 2nd ed.
American Phytopathological Society, St. Paul.
Khan, M.F.R., C.E. Windels, and C.A. Bradley.
2018. Comparison of Cercospora and bacterial
leaf spots on sugar beet. North Dakota State Uni-
versity Extension Service. https://www.ag.ndsu.
edu/publications/crops/comparison-of-cercospora-
and-bacterial-leafspots-on-sugar-beet. [accessed
1 Sep 2021].
Koike, S.T., D.M. Henderson, C.T. Bull, P.H.
Goldman, and R.T. Lewellen. 2003. First report
of bacterial leaf spot of Swiss Chard caused by
Pseudomonas syringae pv. aptata in California.
Plant Dis. 87(11):1397, https://doi.org/10.1094/
pdis.2003.87.11.1397b.
Lamichhane, J.R., A. Messean, and C.E. Morris.
2015. Insights into epidemiology and con-
trol of diseases of annual plants caused by
the Pseudomonas syringae species complex.
J. Gen. Plant Pathol. 81(5):331–350, https://
doi.org/10.1007/s10327-015-0605-z.
May, C., C. Luce, J. Casola, M. Chang, J. Cuha-
ciyan, M. Dalton, S. Lowe, G. Morishima, P.
Mote, A. Petersen, G. Roesch-McNally, and E.
York. 2018. Northwest, p. 1036–1100. In: D.R.
Reidmiller, C.W. Avery, D.R. Easterling, K.E.
Kunkel, K.L.M. Lewis, T.K. Maycock, and
B.C. Stewart (eds.). Impacts, risks, and
1446
adaptation in the United States: Fourth national
climate assessment, Volume II. U.S. Global
Change Research Program, Washington, DC,
https://doi.org/10.7930/NCA4.2018.CH24.
Morris, C.E., C.L. Monteil, and O. Berge. 2013.
The life history of Pseudomonas syringae:
Linking agriculture to earth system processes.
Annu. Rev. Phytopathol. 51:85–104, https://
doi.org/10.1146/annurev-phyto-082712-102402.
Nampijja, M., M. Derie, and L.J. du Toit. 2021.
First report of bacterial leaf spot caused by
Pseudomonas syringae pv. aptata on Swiss
chard, Beta vulgaris subsp. vulgaris, in Ari-
zona. Plant Dis. 105(11):3738, https://doi.org/
10.1094/pdis-12-20-2554-pdn.
Nikolic, I., S. Stankovic, I. Dimkic, T. Beric, V.
Stojsin, J. Janse, and T. Popovic. 2018. Ge-
netic diversity and pathogenicity of Pseudomo-
nas syringae pv. aptata isolated from sugar
beet. Plant Pathol. 67(5):1194–1207, https://
doi.org/10.1111/ppa.12831.
Okazaki, K., N. Ogata, and M. Tanaka. 2005. De-
velopment of zoospore inoculation method for
assay of black root rot resistance in sugar beet.
Jpn. J. Crop. Sci. 74(1):47–51, https://doi.org/
10.1626/jcs.74.47.
Peck, N.H., B.E. Clark, and J.J. Natti. 1967. Effect
of decortication and fungicide on performance
of table beet seed. Agron. J. 59(1):78–80, https://
doi.org/10.2134/agronj1967.00021962005900010
024x.
Pethybridge, S.J., J.R. Kikkert, L.E. Hanson, and
S.C. Nelson. 2018. Challenges and prospects
for building resilient disease management strat-
egies and tactics for the New York table beet
industry. Agronomy (Basel) 8(7):1–17, https://
doi.org/10.3390/agronomy8070112.
Pethybridge, S.J., S. Sharma, Z. Hansen, J.R.
Kikkert, D.L. Olmstead, and L.E. Hanson. 2020.
Optimizing Cercospora leaf spot control in table
beet using action thresholds and disease forecast-
ing. Plant Dis. 104(6):1831–1840, https://doi.org/
10.1094/PDIS-02-20-0246-RE.
Pethybridge, S.J., N. Vaghefi, and J.R. Kikkert. 2017.
Horticultural characteristics and susceptibility of ta-
ble beet cultivars to Cercospora leaf spot in New
York. HortTechnology 27(4):530–538, https://doi.
org/10.21273/HORTTECH03743-17.
Pryor, S.C., D. Scavia, C. Downer, M. Gaden,
L. Iverson, R. Nordstrom, J. Patz, and G.P.
Robertson. 2014. Midwest, p. 418–440. In:
J.M. Melillo, T.C. Richmond, and G.W.
Yohe (eds.). Climate change impacts in the
United States: The third national climate as-
sessment. U.S. Global Change Research Pro-
gram, https://doi.org/10.7930/J0J1012N.
Riffaud, C.M.H. and C.E. Morris. 2002. Detec-
tion of Pseudomonas syringae pv. aptata in
irrigation water retention basins by immuno-
fluorescence colony-staining. Eur. J. Plant
Pathol. 108:539–545, https://doi.org/10.1023/
A:1019919627886.
Rotondo, F., C.M. Vrisman, R. Rani, A.L. Testen,
L. Deblais, and S.A. Miller. 2020. First report
of Pseudomonas syringae pv. aptata causing
bacterial leaf spot on common beet (Beta vul-
garis) in Ohio. Plant Dis. 104(2):561, https://
doi.org/10.1094/pdis-08-19-1720-pdn.
Safni, I., L.R. Sepulveda, P.H. Goldman, M.L. De-
rie, L.J. du Toit, S.T. Koike, V.O. Stockwell,
and C.T. Bull. 2016. Genetic diversity of Pseu-
domonas syringae causing bacterial leaf spot
on table beet (Beta vulgaris) and Swiss chard
(Beta vulgaris subsp. cicla). Phytopathology
106(Suppl 4):143.
Scholten, O.E., L.W. Panella, T.S.M. de Bock, and
W. Lange. 2001. A greenhouse test for screening
sugar beet (Beta vulgaris) for resistance to Rhizoc-
tonia solani. Eur. J. Plant Pathol. 107(2):161–166,
https://doi.org/10.1023/A:1011208903344.
Singh, D. and S.B. Mathur. 2004. Histopathology
and seed-borne infection. CRC Press, Boca Ra-
ton, FL.
U.S. Department of Agriculture. 2019. Census of
agriculture, 2017. https://www.nass.usda.gov/
Publications/AgCensus/2017/index.php#full_
report. [accessed 12 Aug 2022].
Wigg, K.S. and I.L. Goldman. 2020. Variability in re-
action to root and crown rot caused by Rhizoctonia
solani among table beet cultivars, breeding lines,
and plant introductions in controlled environment
conditions. HortScience 55(9):1482–1494, https://
doi.org/10.21273/HORTSCI15011-20.
Wintermantel, W.M. and S.R. Kaffka. 2006. Sugar
beet performance with curly top is related
to virus accumulation and age at infection.
Plant Dis. 90(5):657–662, https://doi.org/10.1094/
PD-90-0657.
Xin, X.F., B. Kvitko, and S.Y. He. 2018. Pseudo-
monas syringae: What it takes to be a pathogen.
Nat. Rev. Microbiol. 16(5):316–328, https://doi.
org/10.1038/nrmicro.2018.17.
HORTSCIENCE VOL. 57(11) NOVEMBER 2022
Supplemental Fig. 1. Mean area under the disease progress curve (AUDPC) of eight Beta vulgaris subsp. vulgaris commercial cultivars, one Beta vulgaris
subsp. vulgaris breeding line, and one Beta vulgaris subsp. cicla commercial cultivar during vegetative growth in a greenhouse screen evaluated in 2021.
Inoculated plants were spray inoculated with PSA strain BP 1452, and control plants were sprayed with sterilized with Milli-Q water.

HORTSCIENCE VOL. 57(11) NOVEMBER 2022 1

Supplemental Fig. 2. Mean area under the disease progress curve (AUDPC) of six Beta vulgaris subsp. vulgaris commercial cultivars, four Beta vulgaris
subsp. vulgaris breeding lines, and six Beta vulgaris subsp. cicla commercial cultivars during vegetative growth in a greenhouse screen evaluated in
2021. Inoculated plants were spray inoculated with PSA strain BP 1452, and control plants were sprayed with sterilized with Milli-Q water.

2 HORTSCIENCE VOL. 57(11) NOVEMBER 2022

Supplemental Fig. 3. Mean area under the disease progress curve (AUDPC) of 26 Beta vulgaris subsp. vulgaris plant introductions (PIs) during vegetative
growth in a greenhouse screen evaluated in 2021. Inoculated plants were spray inoculated with PSA strain BP 1452, and control plants were sprayed with
sterilized with Milli-Q water.

HORTSCIENCE VOL. 57(11) NOVEMBER 2022 3

Supplemental Fig. 4. Mean area under the disease progress curve (AUDPC) of eight Beta vulgaris subsp. vulgaris commercial cultivars, one Beta vulgaris
subsp. vulgaris breeding line, and one Beta vulgaris subsp. cicla commercial cultivar during reproductive growth in a greenhouse screen evaluated in
2021. Inoculated plants were spray inoculated with PSA strain BP 1452, and control plants were sprayed with sterilized with Milli-Q water.

4 HORTSCIENCE VOL. 57(11) NOVEMBER 2022

Supplemental Fig. 5. Mean disease score over days post inoculation of eight Beta vulgaris subsp. vulgaris commercial cultivars, one Beta vulgaris subsp. vul-
garis breeding line, and one Beta vulgaris subsp. cicla commercial cultivar in response to spray inoculation with PSA strain BP 1452 during vegetative
growth in a greenhouse screen evaluated in 2021. The experimental unit for each accession was four inoculated plants and each accession was screened
twice.

Supplemental Fig. 6. Mean disease score over days post inoculation of six Beta vulgaris subsp. vulgaris commercial cultivars, four Beta vulgaris subsp. vul-
garis breeding lines, and six Beta vulgaris subsp. cicla commercial cultivars in response to spray inoculation with PSA strain BP 1452 during vegetative
growth in a greenhouse screen evaluated in 2021. The experimental unit for each accession is four plants. ** refers to significant differences among ac-
cessions at a particular rating time at P <0.05.

HORTSCIENCE VOL. 57(11) NOVEMBER 2022 5

Supplemental Fig. 7. Mean disease score over days post inoculation of 26 Beta vulgaris subsp. vulgaris plant introductions (PIs) in response to spray inocula-
tion with PSA strain BP 1452 during vegetative growth in a greenhouse screen evaluated in 2021. The experimental unit for each accession is four plants.
* refers to significant differences among accessions at a particular rating time at P < 0.1. ** refers to significant differences among accessions at a partic-
ular rating time at P < 0.05.

Supplemental Fig. 8. Mean disease score over days post inoculation of eight Beta vulgaris subsp. vulgaris commercial cultivars, one Beta vulgaris subsp. vul-
garis breeding line, and one Beta vulgaris subsp. cicla commercial cultivar in response to spray inoculation with PSA strain BP 1452 during reproductive
growth in a greenhouse screen evaluated in 2021. The experimental unit for each accession is four plants.

6 HORTSCIENCE VOL. 57(11) NOVEMBER 2022

Supplemental Fig. 9. Difference in appearance of lesions on dark-colored leaves (A) and light-colored leaves (B).

HORTSCIENCE VOL. 57(11) NOVEMBER 2022 7