Neoplastic progression in crown gall in tobacco without elevated auxin levels.

Abstract:

:We have isolated two stable variants from a crown-gall teratoma tissue of tobacco (Nicotiana tabacum L.) transformed by Agrobacterium tumefaciens strain A66, a mutant of the virulent A6 strain containing an insertion sequence in the tumor-inducing (Ti) plasmid at the locus coding for auxin biosynthesis. Normally tobacco cells transformed by strain A66 spontaneously form shoots in culture and will not grow on hormone-free medium unless shoots develop. The variant tissue lines, isolated from the teratoma tissue after prolonged culture in the dark, grew as friable and unorganized tissues on hormone-free growth medium. Growth of the variants was more sensitive to auxin feeding than growth of the parental teratoma line, and the auxin dose-response curves of the variant lines were similar to those obtained with A6-transformed tobacco cells. Southern blot analysis of DNA from the parental teratoma line and one of the variants showed no differences in copy number or organization of the oncogenic DNA sequence (T-DNA) transferred from the bacterium, indicating that the variant phenotype did not result from reversion of the A66 mutation. Radio-immunoassay analysis showed similar levels of indole-3-acetic acid (IAA) in the variants and parental teratoma line (3-50 and 38-42 pmol·(gFW)(-1), respectively), whereas an A6-transformed cell line contained much higher IAA levels (150-1200 pmol·(g FW)(-1)). Low levels of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid in the variants and the parental teratoma line (<5 nmol·(g FW)(-1)) as compared with that found in the A6-transformed line (>100 nmol· (g FW)(-1)) provided additional, indirect evidence for low auxin levels in the variant lines. These results indicate that crown-gall teratoma tissues of tobacco may switch to the unorganized, auxin-sensitive phenotype without an increase in auxin content.

journal_name

Planta

journal_title

Planta

authors

Pengelly WL,Vijayaraghavan SJ,Sciaky D

doi

10.1007/BF00392144

subject

Has Abstract

pub_date

1986-11-01 00:00:00

pages

454-61

issue

3

eissn

0032-0935

issn

1432-2048

journal_volume

169

pub_type

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