Optimizing a protocol for sterilization and in vitro establishment of vegetative buds from mature douglas fir trees

Abdoulaye Traore, Zizhuo Xing, Amy Bonser, John Edward Carlson

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

A successful tissue culture initiation step often begins with effective explant sterilization. To improve douglas fir bud culture initiation, five sterilization treatments (20% bleach, 100% bleach, 3 second flaming, 5 second flaming, and self-extinguishing flaming) were evaluated for their effectiveness on winter and spring bud sterilization. The 20% and 100% bleach treatments resulted in the highest percentage of healthy bud cultures (>90% for winter buds). Spring buds showed a higher level of contamination with 20% bleach sterilization (36%) than did winter buds (1%). Successful sterilisation was also achieved by flaming, but bud injury was observed. Increased flaming time caused a decrease in the percentage of healthy actively growing buds. The percentage of healthy bud cultures after 3 second flaming, 5 second flaming and self-extinguishing-flaming (9 to 14 s) were 66%, 59%, and 10% respectively. In addition, sterilization by either approach required subsequent bud dissection to remove the outer scales; otherwise most buds were lost to contamination. When sterilization was followed by bud dissection, contamination rates for winter buds were <2% for all treatments. After successful sterilization and culture initiation, bud expansion was the highest (50% to 98%) in the presence of low concentrations of BA (0 to 0.045 μmol·L-1), while high concentrations of BA (10.448 to 4.527 μmol·L-1) reduced bud expansion (0% to 60%), but promoted bud multiplication.

Original languageEnglish (US)
Pages (from-to)1464-1468
Number of pages5
JournalHortScience
Volume40
Issue number5
StatePublished - Aug 1 2005

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Pseudotsuga menziesii
buds
winter
bud initiation
tissue culture
explants

All Science Journal Classification (ASJC) codes

  • Horticulture

Cite this

@article{8b339d71145e48858499199836d25c87,
title = "Optimizing a protocol for sterilization and in vitro establishment of vegetative buds from mature douglas fir trees",
abstract = "A successful tissue culture initiation step often begins with effective explant sterilization. To improve douglas fir bud culture initiation, five sterilization treatments (20{\%} bleach, 100{\%} bleach, 3 second flaming, 5 second flaming, and self-extinguishing flaming) were evaluated for their effectiveness on winter and spring bud sterilization. The 20{\%} and 100{\%} bleach treatments resulted in the highest percentage of healthy bud cultures (>90{\%} for winter buds). Spring buds showed a higher level of contamination with 20{\%} bleach sterilization (36{\%}) than did winter buds (1{\%}). Successful sterilisation was also achieved by flaming, but bud injury was observed. Increased flaming time caused a decrease in the percentage of healthy actively growing buds. The percentage of healthy bud cultures after 3 second flaming, 5 second flaming and self-extinguishing-flaming (9 to 14 s) were 66{\%}, 59{\%}, and 10{\%} respectively. In addition, sterilization by either approach required subsequent bud dissection to remove the outer scales; otherwise most buds were lost to contamination. When sterilization was followed by bud dissection, contamination rates for winter buds were <2{\%} for all treatments. After successful sterilization and culture initiation, bud expansion was the highest (50{\%} to 98{\%}) in the presence of low concentrations of BA (0 to 0.045 μmol·L-1), while high concentrations of BA (10.448 to 4.527 μmol·L-1) reduced bud expansion (0{\%} to 60{\%}), but promoted bud multiplication.",
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Optimizing a protocol for sterilization and in vitro establishment of vegetative buds from mature douglas fir trees. / Traore, Abdoulaye; Xing, Zizhuo; Bonser, Amy; Carlson, John Edward.

In: HortScience, Vol. 40, No. 5, 01.08.2005, p. 1464-1468.

Research output: Contribution to journalArticle

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