Studies on Agrobacterium tumefaciens. V. Fate of exogenously added bacterial DNA in Nicotiana tabacum

C. I. Kado; P. F. Lurquin

doi:10.1016/0048-4059(76)90009-6

Studies on Agrobacterium tumefaciens. V. Fate of exogenously added bacterial DNA in Nicotiana tabacum

C. I. Kado, P. F. Lurquin

Research output › peer-review

Abstract

DNA.DNA filter hybridization, DNA solution reassociation enrichment and DNA bouyant density analyses in cesium chloride were used to detect Agrobacterium tumefaciens DNA sequences in exponentially growing tobacco cells that were exposed to the bacterial DNA in axenic culture. The bacterial DNA sequences were not detected in DNA obtained from tobacco callus cells treated with radioactive A. tumefaciens DNA after 4 days growth at 27 °C. Furthermore, no radioactive DNA with a bouyant density intermediate between tobacco DNA and bacterial donor DNA was observed. Ultrasonication of DNA from DNA-treated callus cells did not release any presumptive A. tumefaciens DNA. Moreover, A. tumefaciens DNA was not replicated but degraded and re-utilized by the callus cells within the 4 days of incubation. Any traces of presumptive replicated bacterial DNA (< 0·01% of the total callus DNA) were enriched and separated as double-stranded DNA by hydroxylapatite chromatography. The thermal stabilities, melting characteristics and T_m of this enriched DNA were identical to that of enriched DNA derived from untreated callus cell DNA and clearly distinct from that of the reassociated A. tumefaciens DNA. These results are in contrast to previous reports of uptake, integration and replication of bacterial DNA in higher plants, and do not lend supporting evidence for direct transformation of callus cells in axenic culture by the addition of purified A. tumefaciens DNA.

Original language	English
Pages (from-to)	73-82
Number of pages	10
Journal	Physiological Plant Pathology
Volume	8
Issue number	1
DOIs	https://doi.org/10.1016/0048-4059(76)90009-6
State	Published - Jan 1976

Funding

This research was supported by NIH grant CA-I 1526 from the National Cancer Institute.

Funders	Funder number
National Cancer Institute (NCI)	CA-I 1526

Access to Document

10.1016/0048-4059(76)90009-6

Cite this

@article{4c1cd0f41cdb43bd80d5a3bf29460add,

title = "Studies on Agrobacterium tumefaciens. V. Fate of exogenously added bacterial DNA in Nicotiana tabacum",

abstract = "DNA.DNA filter hybridization, DNA solution reassociation enrichment and DNA bouyant density analyses in cesium chloride were used to detect Agrobacterium tumefaciens DNA sequences in exponentially growing tobacco cells that were exposed to the bacterial DNA in axenic culture. The bacterial DNA sequences were not detected in DNA obtained from tobacco callus cells treated with radioactive A. tumefaciens DNA after 4 days growth at 27 °C. Furthermore, no radioactive DNA with a bouyant density intermediate between tobacco DNA and bacterial donor DNA was observed. Ultrasonication of DNA from DNA-treated callus cells did not release any presumptive A. tumefaciens DNA. Moreover, A. tumefaciens DNA was not replicated but degraded and re-utilized by the callus cells within the 4 days of incubation. Any traces of presumptive replicated bacterial DNA (< 0·01% of the total callus DNA) were enriched and separated as double-stranded DNA by hydroxylapatite chromatography. The thermal stabilities, melting characteristics and Tm of this enriched DNA were identical to that of enriched DNA derived from untreated callus cell DNA and clearly distinct from that of the reassociated A. tumefaciens DNA. These results are in contrast to previous reports of uptake, integration and replication of bacterial DNA in higher plants, and do not lend supporting evidence for direct transformation of callus cells in axenic culture by the addition of purified A. tumefaciens DNA.",

author = "Kado, {C. I.} and Lurquin, {P. F.}",

year = "1976",

month = jan,

doi = "10.1016/0048-4059(76)90009-6",

language = "English",

volume = "8",

pages = "73--82",

number = "1",

}

TY - JOUR

T1 - Studies on Agrobacterium tumefaciens. V. Fate of exogenously added bacterial DNA in Nicotiana tabacum

AU - Kado, C. I.

AU - Lurquin, P. F.

PY - 1976/1

Y1 - 1976/1

N2 - DNA.DNA filter hybridization, DNA solution reassociation enrichment and DNA bouyant density analyses in cesium chloride were used to detect Agrobacterium tumefaciens DNA sequences in exponentially growing tobacco cells that were exposed to the bacterial DNA in axenic culture. The bacterial DNA sequences were not detected in DNA obtained from tobacco callus cells treated with radioactive A. tumefaciens DNA after 4 days growth at 27 °C. Furthermore, no radioactive DNA with a bouyant density intermediate between tobacco DNA and bacterial donor DNA was observed. Ultrasonication of DNA from DNA-treated callus cells did not release any presumptive A. tumefaciens DNA. Moreover, A. tumefaciens DNA was not replicated but degraded and re-utilized by the callus cells within the 4 days of incubation. Any traces of presumptive replicated bacterial DNA (< 0·01% of the total callus DNA) were enriched and separated as double-stranded DNA by hydroxylapatite chromatography. The thermal stabilities, melting characteristics and Tm of this enriched DNA were identical to that of enriched DNA derived from untreated callus cell DNA and clearly distinct from that of the reassociated A. tumefaciens DNA. These results are in contrast to previous reports of uptake, integration and replication of bacterial DNA in higher plants, and do not lend supporting evidence for direct transformation of callus cells in axenic culture by the addition of purified A. tumefaciens DNA.

AB - DNA.DNA filter hybridization, DNA solution reassociation enrichment and DNA bouyant density analyses in cesium chloride were used to detect Agrobacterium tumefaciens DNA sequences in exponentially growing tobacco cells that were exposed to the bacterial DNA in axenic culture. The bacterial DNA sequences were not detected in DNA obtained from tobacco callus cells treated with radioactive A. tumefaciens DNA after 4 days growth at 27 °C. Furthermore, no radioactive DNA with a bouyant density intermediate between tobacco DNA and bacterial donor DNA was observed. Ultrasonication of DNA from DNA-treated callus cells did not release any presumptive A. tumefaciens DNA. Moreover, A. tumefaciens DNA was not replicated but degraded and re-utilized by the callus cells within the 4 days of incubation. Any traces of presumptive replicated bacterial DNA (< 0·01% of the total callus DNA) were enriched and separated as double-stranded DNA by hydroxylapatite chromatography. The thermal stabilities, melting characteristics and Tm of this enriched DNA were identical to that of enriched DNA derived from untreated callus cell DNA and clearly distinct from that of the reassociated A. tumefaciens DNA. These results are in contrast to previous reports of uptake, integration and replication of bacterial DNA in higher plants, and do not lend supporting evidence for direct transformation of callus cells in axenic culture by the addition of purified A. tumefaciens DNA.

UR - http://www.scopus.com/inward/record.url?scp=49549133731&partnerID=8YFLogxK

U2 - 10.1016/0048-4059(76)90009-6

DO - 10.1016/0048-4059(76)90009-6

M3 - Article

AN - SCOPUS:49549133731

SN - 0048-4059

VL - 8

SP - 73

EP - 82

JO - Physiological Plant Pathology

JF - Physiological Plant Pathology

IS - 1

ER -