Transient reporter gene (GUS) expression in creeping bentgrass (Agrostis palustris) is affected by in vivo nucleolytic activity.
Leaf and callus tissues of a creeping bentgrass cultivar (Penn A4) had high nuclease activities that degraded exogenously added plasmid DNA. When callus tissue was incubated for 24 h with heparin, spermidine, aurintricarboxylic acid or polyethylene glycol, only heparin and spermidine were effective as in vitro nuclease inhibitors, protecting exogenously added plasmid DNA from degradation. When beta-glucuronidase (GUS) reporter gene activity was evaluated in heparin-treated (0.6%), 14-month old callus following microprojectile bombardment, GUS activity increased 1000-fold compared to equivalent aged untreated Penn A4 callus. Similar enhancement from heparin pretreatment (0.6% or 1.2%) was not observed in 6-month old callus. This is likely due to much higher activities of nuclease in the younger callus.
Basu
Chhandak
C
Department of Plant Sciences, University of Rhode Island, 9 East Alumni Avenue, Suite 7, Kingston, Rhode Island 02881, USA.
Luo
Hong
H
Kausch
Albert P
AP
Chandlee
Joel M
JM
D000375:Aging / Q000502:physiology; D031703:Agrostis / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D001312:Aurintricarboxylic Acid / Q000494:pharmacology; D046508:Culture Techniques; D003851:Deoxyribonucleases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter / Q000235:genetics; D005966:Glucuronidase / Q000096:biosynthesis* / Q000235:genetics*; D006493:Heparin / Q000494:pharmacology; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000008:administration & dosage; D011092:Polyethylene Glycols / Q000494:pharmacology; D013045:Species Specificity; D013095:Spermidine / Q000494:pharmacology; D014170:Transformation, Genetic / Q000235:genetics; D019076:Transgenes / Q000235:genetics
D011092:Polyethylene Glycols; D001312:Aurintricarboxylic Acid; D006493:Heparin; D003851:Deoxyribonucleases; D005966:Glucuronidase; D013095:Spermidine