Colonization of siliques and seeds of rapid cycling Brassica oleracea plants by Xanthomonas campestris pv. campestris after spray-inoculation of flower clusters.

J.M. van der Wolf, P. Kastelein, Tadeu Antônio Fernandes da Silva Júnior, Flavia Vieira Lelis, P.S. van der Zouwen

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Glasshouse experiments were conducted to study the colonization of seedpods (siliques) and seeds of rapid cycling Brassica oleracea plants after spraying
inoculum on clusters of recently opened flowers with Xanthomonas campestris pv. campestris (Xcc) at densities of 107–108 cfu ml−1. A green fluorescent protein (GFP) tagged Xcc strain was used to allow visualization of the bacteria by epifluorescence stereo microscopy (ESM) and confocal laser scanning microscopy (CLSM). The GFP-tagged strain showed reduced virulence compared to the untagged parental strain, but was still able to cause black rot symptoms. Two to three days after sprayinoculation, sepals, stamen and petals were colonized by
Xcc, as observed by ESM. In green siliques a GFP-signal was observed on valves, septa and seeds, despite the fact that a high percentage of Xcc cells had lost their ability to express GFP as found by dilution-plating. Densities of Xcc in infected silique tissues were up to 109 cfu g−1. A fluorescent signal using ESM was found in seeds harvested from symptomatic siliques after incubation of
seeds on blotting paper wetted with broth to enhance the multiplication of Xcc. Xcc was found in association with the seed coat and in a single seed, also in the
endosperm and embryo, indicating deep-seated seed infection. The estimated incidence of contaminated seeds in both years was ca. 7%. The estimated incidence of deep-seated infections, still detectable after warm water treatment of seeds, was also high (2–3.8%). It is concluded that spray-inoculation of flower clusters with Xcc can result in the infection of sepals and reproductive organs,
and in deep-seated seed infections.
LanguageEnglish
Pages445-461
Number of pages17
JournalEuropean Journal of Plant Pathology
Volume154
Issue number2
Early online date4 Feb 2019
DOIs
Publication statusPublished - Jun 2019

Fingerprint

Xanthomonas campestris pv. campestris
Brassica oleracea
flowers
seeds
green fluorescent protein
microscopy
calyx
infection
incidence
valves (equipment)
confocal laser scanning microscopy
stamens
water treatment
embryo (plant)
signs and symptoms (plants)
corolla
virulence
greenhouses

Cite this

@article{488c6c3a069f4b2eabc9e3f865f8045c,
title = "Colonization of siliques and seeds of rapid cycling Brassica oleracea plants by Xanthomonas campestris pv. campestris after spray-inoculation of flower clusters.",
abstract = "Glasshouse experiments were conducted to study the colonization of seedpods (siliques) and seeds of rapid cycling Brassica oleracea plants after sprayinginoculum on clusters of recently opened flowers with Xanthomonas campestris pv. campestris (Xcc) at densities of 107–108 cfu ml−1. A green fluorescent protein (GFP) tagged Xcc strain was used to allow visualization of the bacteria by epifluorescence stereo microscopy (ESM) and confocal laser scanning microscopy (CLSM). The GFP-tagged strain showed reduced virulence compared to the untagged parental strain, but was still able to cause black rot symptoms. Two to three days after sprayinoculation, sepals, stamen and petals were colonized byXcc, as observed by ESM. In green siliques a GFP-signal was observed on valves, septa and seeds, despite the fact that a high percentage of Xcc cells had lost their ability to express GFP as found by dilution-plating. Densities of Xcc in infected silique tissues were up to 109 cfu g−1. A fluorescent signal using ESM was found in seeds harvested from symptomatic siliques after incubation ofseeds on blotting paper wetted with broth to enhance the multiplication of Xcc. Xcc was found in association with the seed coat and in a single seed, also in theendosperm and embryo, indicating deep-seated seed infection. The estimated incidence of contaminated seeds in both years was ca. 7{\%}. The estimated incidence of deep-seated infections, still detectable after warm water treatment of seeds, was also high (2–3.8{\%}). It is concluded that spray-inoculation of flower clusters with Xcc can result in the infection of sepals and reproductive organs,and in deep-seated seed infections.",
author = "{van der Wolf}, J.M. and P. Kastelein and {Ant{\^o}nio Fernandes da Silva J{\'u}nior}, Tadeu and {Vieira Lelis}, Flavia and {van der Zouwen}, P.S.",
year = "2019",
month = "6",
doi = "10.1007/s10658-019-01668-4",
language = "English",
volume = "154",
pages = "445--461",
journal = "European Journal of Plant Pathology",
issn = "0929-1873",
publisher = "Springer Verlag",
number = "2",

}

Colonization of siliques and seeds of rapid cycling Brassica oleracea plants by Xanthomonas campestris pv. campestris after spray-inoculation of flower clusters. / van der Wolf, J.M.; Kastelein, P.; Antônio Fernandes da Silva Júnior, Tadeu; Vieira Lelis, Flavia; van der Zouwen, P.S.

In: European Journal of Plant Pathology, Vol. 154, No. 2, 06.2019, p. 445-461.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Colonization of siliques and seeds of rapid cycling Brassica oleracea plants by Xanthomonas campestris pv. campestris after spray-inoculation of flower clusters.

AU - van der Wolf, J.M.

AU - Kastelein, P.

AU - Antônio Fernandes da Silva Júnior, Tadeu

AU - Vieira Lelis, Flavia

AU - van der Zouwen, P.S.

PY - 2019/6

Y1 - 2019/6

N2 - Glasshouse experiments were conducted to study the colonization of seedpods (siliques) and seeds of rapid cycling Brassica oleracea plants after sprayinginoculum on clusters of recently opened flowers with Xanthomonas campestris pv. campestris (Xcc) at densities of 107–108 cfu ml−1. A green fluorescent protein (GFP) tagged Xcc strain was used to allow visualization of the bacteria by epifluorescence stereo microscopy (ESM) and confocal laser scanning microscopy (CLSM). The GFP-tagged strain showed reduced virulence compared to the untagged parental strain, but was still able to cause black rot symptoms. Two to three days after sprayinoculation, sepals, stamen and petals were colonized byXcc, as observed by ESM. In green siliques a GFP-signal was observed on valves, septa and seeds, despite the fact that a high percentage of Xcc cells had lost their ability to express GFP as found by dilution-plating. Densities of Xcc in infected silique tissues were up to 109 cfu g−1. A fluorescent signal using ESM was found in seeds harvested from symptomatic siliques after incubation ofseeds on blotting paper wetted with broth to enhance the multiplication of Xcc. Xcc was found in association with the seed coat and in a single seed, also in theendosperm and embryo, indicating deep-seated seed infection. The estimated incidence of contaminated seeds in both years was ca. 7%. The estimated incidence of deep-seated infections, still detectable after warm water treatment of seeds, was also high (2–3.8%). It is concluded that spray-inoculation of flower clusters with Xcc can result in the infection of sepals and reproductive organs,and in deep-seated seed infections.

AB - Glasshouse experiments were conducted to study the colonization of seedpods (siliques) and seeds of rapid cycling Brassica oleracea plants after sprayinginoculum on clusters of recently opened flowers with Xanthomonas campestris pv. campestris (Xcc) at densities of 107–108 cfu ml−1. A green fluorescent protein (GFP) tagged Xcc strain was used to allow visualization of the bacteria by epifluorescence stereo microscopy (ESM) and confocal laser scanning microscopy (CLSM). The GFP-tagged strain showed reduced virulence compared to the untagged parental strain, but was still able to cause black rot symptoms. Two to three days after sprayinoculation, sepals, stamen and petals were colonized byXcc, as observed by ESM. In green siliques a GFP-signal was observed on valves, septa and seeds, despite the fact that a high percentage of Xcc cells had lost their ability to express GFP as found by dilution-plating. Densities of Xcc in infected silique tissues were up to 109 cfu g−1. A fluorescent signal using ESM was found in seeds harvested from symptomatic siliques after incubation ofseeds on blotting paper wetted with broth to enhance the multiplication of Xcc. Xcc was found in association with the seed coat and in a single seed, also in theendosperm and embryo, indicating deep-seated seed infection. The estimated incidence of contaminated seeds in both years was ca. 7%. The estimated incidence of deep-seated infections, still detectable after warm water treatment of seeds, was also high (2–3.8%). It is concluded that spray-inoculation of flower clusters with Xcc can result in the infection of sepals and reproductive organs,and in deep-seated seed infections.

U2 - 10.1007/s10658-019-01668-4

DO - 10.1007/s10658-019-01668-4

M3 - Article

VL - 154

SP - 445

EP - 461

JO - European Journal of Plant Pathology

T2 - European Journal of Plant Pathology

JF - European Journal of Plant Pathology

SN - 0929-1873

IS - 2

ER -