TY - JOUR
T1 - Optimizing a protocol for sterilization and in vitro establishment of vegetative buds from mature douglas fir trees
AU - Traore, Abdoulaye
AU - Xing, Zizhuo
AU - Bonser, Amy
AU - Carlson, John
PY - 2005/8
Y1 - 2005/8
N2 - 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.
AB - 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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U2 - 10.21273/hortsci.40.5.1464
DO - 10.21273/hortsci.40.5.1464
M3 - Article
AN - SCOPUS:22444446665
SN - 0018-5345
VL - 40
SP - 1464
EP - 1468
JO - HortScience
JF - HortScience
IS - 5
ER -