Isolation and characterisation of a pod dehiscence zone-specific polygalacturonase from Brassica napus.
Seven distinct partial cDNAs, similar in sequence to previously described polygalacturonases (PGs), were amplified from cDNA derived from rape pod wall, dehiscence zone and leaves by the polymerase chain reaction. Northern analysis showed that one clone, PG35-8, was expressed at low levels in the dehiscence zone during the first five weeks after anthesis but was very abundantly expressed at week 6. In contrast, no PG35-8-related RNA was detected in the pod wall. Our data suggest that there are temporal and spatial correlations between the breakdown of the middle lamella, of the dehiscence zone cells and the pattern of synthesis of PG35-8 transcripts which may indicate a role for this particular PG in rape pod dehiscence. PG35-8 was used to isolate five cDNA clones from a rape dehiscence zone cDNA library. Restriction enzyme analysis and partial sequencing revealed that they were derived from four highly homologous transcripts which are probably allelic forms of a single gene. One full-length clone, RDPG1, was completely sequenced. The predicted protein of RDPG1 showed its highest identity with PG from apple fruit with an identity of 52%.
Petersen
M
M
Biotechnology Group, DIPS, Lyngby.
Sander
L
L
Child
R
R
van Onckelen
H
H
Ulvskov
P
P
Borkhardt
B
B
D000595:Amino Acid Sequence; D001483:Base Sequence; D001937:Brassica / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D002473:Cell Wall / Q000201:enzymology; D017931:DNA Primers; D018076:DNA, Complementary; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D015388:Organelles / Q000502:physiology / Q000648:ultrastructure; D010944:Plants / Q000201:enzymology; D011096:Polygalacturonase / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D018076:DNA, Complementary; D011096:Polygalacturonase