MOLECULAR GENETIC-ANALYSIS OF CHALCONE SYNTHASE IN LYCOPERSICON-ESCULENTUM AND AN ANTHOCYANIN-DEFICIENT MUTANT

被引:63
作者
ONEILL, SD [1 ]
TONG, Y [1 ]
SPORLEIN, B [1 ]
FORKMANN, G [1 ]
YODER, JI [1 ]
机构
[1] UNIV CALIF DAVIS,DEPT VEGETABLE CROPS,DAVIS,CA 95616
来源
MOLECULAR & GENERAL GENETICS | 1990年 / 224卷 / 02期
关键词
Anthocyanin-less mutants; cDNA; Chalcone synthase; Tomato;
D O I
10.1007/BF00271562
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Twelve loci have previously been identified in tomato (Lycopersicon esculentum) that control the intensity and distribution of anthocyanin pigmentation; these are useful genetic markers because they encode phenotypes that are readily visualized in the hypocotyls of emerging seedlings. In order to obtain molecular probes for tomato anthocyanin biosynthesis genes, we isolated two cDNAs which encode chalcone synthase (CHS), one of the key enzymes in anthocyanin biosynthesis, from a tomato hypocotyl cDNA library. By comparing their nucleic acid sequences, we determined that the two CHS cDNAs have an overall similarity of 76% at the nucleotide level and 88% at the amino acid level. We identified hybridization conditions that would distinguish the two clones and by Northern analysis showed that 1.5 kb mRNA species corresponding to each cDNA were expressed in cotyledons, hypocotyls and leaves of wild-type seedlings. Hybridization of the cDNAs at low stringency to genomic blots indicated that in tomato, CHS genes comprise a family of at least three individual members. The two genes that encode the CHS cDNAs were then placed onto the tomato genetic map at unique loci by restriction fragment length polymorphism mapping. We also assayed the activity of CHS and another enzyme in the anthocyanin pathway, flavone 3-hydroxylase, in hypocotyl extracts of wild-type tomato and a number of anthocyanin-deficient mutants. Five mutants had reduced CHS activity when compared to the wildtype controls. Of these, three were also reduce in flavone 3-hydroxylase activity, suggesting a regulatory role for these loci. The other two mutants were preferentially reduced in CHS activity, suggesting a more specific role for these loci in CHS expression. © 1990 Springer-Verlag.
引用
收藏
页码:279 / 288
页数:10
相关论文
共 45 条
[31]   CYTOGENETICS OF THE TOMATO [J].
RICK, CM ;
BUTLER, L .
ADVANCES IN GENETICS, 1956, 8 :267-382
[32]  
RICK CM, 1987, TOMATO GENET COOP RE, V37, P92
[33]   LABELING DEOXYRIBONUCLEIC-ACID TO HIGH SPECIFIC ACTIVITY INVITRO BY NICK TRANSLATION WITH DNA-POLYMERASE I [J].
RIGBY, PWJ ;
DIECKMANN, M ;
RHODES, C ;
BERG, P .
JOURNAL OF MOLECULAR BIOLOGY, 1977, 113 (01) :237-251
[34]   ORGANIZATION AND DIFFERENTIAL ACTIVATION OF A GENE FAMILY ENCODING THE PLANT DEFENSE ENZYME CHALCONE SYNTHASE IN PHASEOLUS-VULGARIS [J].
RYDER, TB ;
HEDRICK, SA ;
BELL, JN ;
LIANG, XW ;
CLOUSE, SD ;
LAMB, CJ .
MOLECULAR AND GENERAL GENETICS, 1987, 210 (02) :219-233
[35]   DNA SEQUENCING WITH CHAIN-TERMINATING INHIBITORS [J].
SANGER, F ;
NICKLEN, S ;
COULSON, AR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (12) :5463-5467
[36]   STRUCTURE OF THE CHALCONE SYNTHASE GENE OF ANTIRRHINUM-MAJUS [J].
SOMMER, H ;
SAEDLER, H .
MOLECULAR AND GENERAL GENETICS, 1986, 202 (03) :429-434
[37]   CHALCONE SYNTHESIS AND HYDROXYLATION OF FLAVONOIDS IN 3'-POSITION WITH ENZYME PREPARATIONS FROM FLOWERS OF DIANTHUS-CARYOPHYLLUS L (CARNATION) [J].
SPRIBILLE, R ;
FORKMANN, G .
PLANTA, 1982, 155 (02) :176-182
[38]   LINKAGE-1 - A PASCAL COMPUTER-PROGRAM FOR THE DETECTION AND ANALYSIS OF GENETIC-LINKAGE [J].
SUITER, KA ;
WENDEL, JF ;
CASE, JS .
JOURNAL OF HEREDITY, 1983, 74 (03) :203-204
[39]  
THOMAS PS, 1983, METHOD ENZYMOL, V100, P255
[40]   THE PARAMUTAGENIC LINE NIV-44 HAS A 5 KB INSERT, TAM-2, IN THE CHALCONE SYNTHASE GENE OF ANTIRRHINUM-MAJUS [J].
UPADHYAYA, KC ;
SOMMER, H ;
KREBBERS, E ;
SAEDLER, H .
MOLECULAR & GENERAL GENETICS, 1985, 199 (02) :201-207