A physical, enzymatic, and genetic characterization of perturbations in the seeds of the brownseed tomato mutants

A. Bruce Downie; Lynnette M.A. Dirk; Qilong Xu; Jennifer Drake; Deqing Zhang; Manjul Dutt; Alan Butterfield; Robert R. Geneve; J. Willis Corum; Karl G. Lindstrom; John C. Snyder

doi:10.1093/jxb/erh112

A physical, enzymatic, and genetic characterization of perturbations in the seeds of the brownseed tomato mutants

A. Bruce Downie, Lynnette M.A. Dirk, Qilong Xu, Jennifer Drake, Deqing Zhang, Manjul Dutt, Alan Butterfield, Robert R. Geneve, J. Willis Corum, Karl G. Lindstrom, John C. Snyder

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

The brownseed mutants (bs¹, bs², and bs⁴) of tomato all possess dark testae and deleteriously affect seed germination speed and/or final percentage. Poor germination performance of the bs ¹ but not the bs⁴ mutant, was due to greater impediment to radicle egress. Testa toughening (bs¹) was prevented by drying in N₂. However, poor germination speed was hardly affected by drying. GA₄₊₇ did not ameliorate germination percentage or speed (bs ¹, bs²), whereas bs⁴ seeds commenced radicle protrusion sooner and had a greater germination percentage. bs¹ mutant seeds have two times more catalase activity while those of bs⁴ contained six times more peroxidase and almost two times more catalase activity than WTs. bs⁴ release only half of the reactive oxygen species into the media than WT during imbibition. EPR detected the presence of free radicals in bs¹ and its WT. bs mutants were epistatic to 12 anthocyaninless mutations, at least some of which produce seeds of lighter than usual testa colour. Macro-arrays of subtractive, suppressive PCR products identified differentially regulated transcripts between seeds of bs⁴ and WT. EST Identity suggests bs⁴ does not exit the developmental programme upon attaining maturity.

Original language	English
Pages (from-to)	961-973
Number of pages	13
Journal	Journal of Experimental Botany
Volume	55
Issue number	399
DOIs	https://doi.org/10.1093/jxb/erh112
State	Published - May 2004

Bibliographical note

Funding Information:
The gibberellic acid (GA4+7) used in these experiments was the kind gift of Abbott Biochemicals, Chicago, IL. The lateral suppressor, brownseed, and anthocyaninless mutant lines were provided by Roger Chetelat from stock maintained at the Charles M Rick Tomato Genetics Resource Center, UC Davis, CA, USA. Janet A Pfeiffer expertly maintained the capillary mat bench, the health of the plants, and the greenhouse facility. Mr David McNertney kindly allowed the use of a Paradigm Seedlot Vigor Assessment™ system (version 3.2; Paradigm Research Corp, South Haven, MN, USA) for performing and documenting the mutant evaluations. Daryl Slone, Kay Oakley, and Dave Lowrey maintained the plants at the University of Kentucky Horticulture Experimental Farm. Ms Love Gill (UK-Advanced Genetics Technologies Center, AGTC) performed the plasmid preparation, cycle sequencing reactions, clean-up, and analysis. Mr Venu-Gopal Puram (UK–Kentucky Biomedical Research Infrastructure Network, UK–KBRIN, Department of Biology) oversaw sequence quality, vector/adaptor masking, and batch BLAST analysis using the UK-AGTC data pipeline. This work was supported by the Department of Horticulture, University of Kentucky, Hatch funds, a Kentucky NSF EPSCoR grant, and CSREES/USDA Special Research Grant (2003-34457-13114).

Keywords

Catalase
Electron paramagnetic resonance (EPR)
Free radicals
Germination
Lycopersicon esculentum
Macro-array
Peroxidase
Seed
Subtractive-suppressive PCR
Testa

ASJC Scopus subject areas

Physiology
Plant Science

Access to Document

10.1093/jxb/erh112

Cite this

@article{705b4173e0d2447598535ab2ff757fb1,

title = "A physical, enzymatic, and genetic characterization of perturbations in the seeds of the brownseed tomato mutants",

abstract = "The brownseed mutants (bs1, bs2, and bs4) of tomato all possess dark testae and deleteriously affect seed germination speed and/or final percentage. Poor germination performance of the bs 1 but not the bs4 mutant, was due to greater impediment to radicle egress. Testa toughening (bs1) was prevented by drying in N2. However, poor germination speed was hardly affected by drying. GA4+7 did not ameliorate germination percentage or speed (bs 1, bs2), whereas bs4 seeds commenced radicle protrusion sooner and had a greater germination percentage. bs1 mutant seeds have two times more catalase activity while those of bs4 contained six times more peroxidase and almost two times more catalase activity than WTs. bs4 release only half of the reactive oxygen species into the media than WT during imbibition. EPR detected the presence of free radicals in bs1 and its WT. bs mutants were epistatic to 12 anthocyaninless mutations, at least some of which produce seeds of lighter than usual testa colour. Macro-arrays of subtractive, suppressive PCR products identified differentially regulated transcripts between seeds of bs4 and WT. EST Identity suggests bs4 does not exit the developmental programme upon attaining maturity.",

keywords = "Catalase, Electron paramagnetic resonance (EPR), Free radicals, Germination, Lycopersicon esculentum, Macro-array, Peroxidase, Seed, Subtractive-suppressive PCR, Testa",

author = "Downie, {A. Bruce} and Dirk, {Lynnette M.A.} and Qilong Xu and Jennifer Drake and Deqing Zhang and Manjul Dutt and Alan Butterfield and Geneve, {Robert R.} and Corum, {J. Willis} and Lindstrom, {Karl G.} and Snyder, {John C.}",

note = "Funding Information: The gibberellic acid (GA4+7) used in these experiments was the kind gift of Abbott Biochemicals, Chicago, IL. The lateral suppressor, brownseed, and anthocyaninless mutant lines were provided by Roger Chetelat from stock maintained at the Charles M Rick Tomato Genetics Resource Center, UC Davis, CA, USA. Janet A Pfeiffer expertly maintained the capillary mat bench, the health of the plants, and the greenhouse facility. Mr David McNertney kindly allowed the use of a Paradigm Seedlot Vigor Assessment{\texttrademark} system (version 3.2; Paradigm Research Corp, South Haven, MN, USA) for performing and documenting the mutant evaluations. Daryl Slone, Kay Oakley, and Dave Lowrey maintained the plants at the University of Kentucky Horticulture Experimental Farm. Ms Love Gill (UK-Advanced Genetics Technologies Center, AGTC) performed the plasmid preparation, cycle sequencing reactions, clean-up, and analysis. Mr Venu-Gopal Puram (UK–Kentucky Biomedical Research Infrastructure Network, UK–KBRIN, Department of Biology) oversaw sequence quality, vector/adaptor masking, and batch BLAST analysis using the UK-AGTC data pipeline. This work was supported by the Department of Horticulture, University of Kentucky, Hatch funds, a Kentucky NSF EPSCoR grant, and CSREES/USDA Special Research Grant (2003-34457-13114).",

year = "2004",

month = may,

doi = "10.1093/jxb/erh112",

language = "English",

volume = "55",

pages = "961--973",

number = "399",

}

TY - JOUR

T1 - A physical, enzymatic, and genetic characterization of perturbations in the seeds of the brownseed tomato mutants

AU - Downie, A. Bruce

AU - Dirk, Lynnette M.A.

AU - Xu, Qilong

AU - Drake, Jennifer

AU - Zhang, Deqing

AU - Dutt, Manjul

AU - Butterfield, Alan

AU - Geneve, Robert R.

AU - Corum, J. Willis

AU - Lindstrom, Karl G.

AU - Snyder, John C.

N1 - Funding Information: The gibberellic acid (GA4+7) used in these experiments was the kind gift of Abbott Biochemicals, Chicago, IL. The lateral suppressor, brownseed, and anthocyaninless mutant lines were provided by Roger Chetelat from stock maintained at the Charles M Rick Tomato Genetics Resource Center, UC Davis, CA, USA. Janet A Pfeiffer expertly maintained the capillary mat bench, the health of the plants, and the greenhouse facility. Mr David McNertney kindly allowed the use of a Paradigm Seedlot Vigor Assessment™ system (version 3.2; Paradigm Research Corp, South Haven, MN, USA) for performing and documenting the mutant evaluations. Daryl Slone, Kay Oakley, and Dave Lowrey maintained the plants at the University of Kentucky Horticulture Experimental Farm. Ms Love Gill (UK-Advanced Genetics Technologies Center, AGTC) performed the plasmid preparation, cycle sequencing reactions, clean-up, and analysis. Mr Venu-Gopal Puram (UK–Kentucky Biomedical Research Infrastructure Network, UK–KBRIN, Department of Biology) oversaw sequence quality, vector/adaptor masking, and batch BLAST analysis using the UK-AGTC data pipeline. This work was supported by the Department of Horticulture, University of Kentucky, Hatch funds, a Kentucky NSF EPSCoR grant, and CSREES/USDA Special Research Grant (2003-34457-13114).

PY - 2004/5

Y1 - 2004/5

N2 - The brownseed mutants (bs1, bs2, and bs4) of tomato all possess dark testae and deleteriously affect seed germination speed and/or final percentage. Poor germination performance of the bs 1 but not the bs4 mutant, was due to greater impediment to radicle egress. Testa toughening (bs1) was prevented by drying in N2. However, poor germination speed was hardly affected by drying. GA4+7 did not ameliorate germination percentage or speed (bs 1, bs2), whereas bs4 seeds commenced radicle protrusion sooner and had a greater germination percentage. bs1 mutant seeds have two times more catalase activity while those of bs4 contained six times more peroxidase and almost two times more catalase activity than WTs. bs4 release only half of the reactive oxygen species into the media than WT during imbibition. EPR detected the presence of free radicals in bs1 and its WT. bs mutants were epistatic to 12 anthocyaninless mutations, at least some of which produce seeds of lighter than usual testa colour. Macro-arrays of subtractive, suppressive PCR products identified differentially regulated transcripts between seeds of bs4 and WT. EST Identity suggests bs4 does not exit the developmental programme upon attaining maturity.

AB - The brownseed mutants (bs1, bs2, and bs4) of tomato all possess dark testae and deleteriously affect seed germination speed and/or final percentage. Poor germination performance of the bs 1 but not the bs4 mutant, was due to greater impediment to radicle egress. Testa toughening (bs1) was prevented by drying in N2. However, poor germination speed was hardly affected by drying. GA4+7 did not ameliorate germination percentage or speed (bs 1, bs2), whereas bs4 seeds commenced radicle protrusion sooner and had a greater germination percentage. bs1 mutant seeds have two times more catalase activity while those of bs4 contained six times more peroxidase and almost two times more catalase activity than WTs. bs4 release only half of the reactive oxygen species into the media than WT during imbibition. EPR detected the presence of free radicals in bs1 and its WT. bs mutants were epistatic to 12 anthocyaninless mutations, at least some of which produce seeds of lighter than usual testa colour. Macro-arrays of subtractive, suppressive PCR products identified differentially regulated transcripts between seeds of bs4 and WT. EST Identity suggests bs4 does not exit the developmental programme upon attaining maturity.

KW - Catalase

KW - Electron paramagnetic resonance (EPR)

KW - Free radicals

KW - Germination

KW - Lycopersicon esculentum

KW - Macro-array

KW - Peroxidase

KW - Seed

KW - Subtractive-suppressive PCR

KW - Testa

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U2 - 10.1093/jxb/erh112

DO - 10.1093/jxb/erh112

M3 - Article

C2 - 15073220

AN - SCOPUS:2442485783

SN - 0022-0957

VL - 55

SP - 961

EP - 973

JO - Journal of Experimental Botany

JF - Journal of Experimental Botany

IS - 399

ER -

A physical, enzymatic, and genetic characterization of perturbations in the seeds of the brownseed tomato mutants

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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