Abstract
Suppression of gene expression using antisense technology has been successful in various applications. In this paper we report differential inhibition of gene expression of the chalcone synthase (chs) gene superfamily members in transgenic Gerbera hybrida (Asteraceae) plants. We have transformed two different cDNAs of the chs gene family, gchs 1 [4] and gchs2, in antisense orientation under control of the CaMV 35S promoter into gerbera. Gchs1 codes for an enzyme with chalcone synthase activity while gchs2 is a more diverged member of the gene family having distinct structure and expression pattern. Furthermore, gchs2 is evidently not involved in anthocyanin synthesis and encodes an enzyme with novel catalytic properties. In both cases effective blocking of the resident sense gene expression was detected. In addition, the transformation affected differentially the expression of other members of the chs gene family. The degree of inhibition appeared to depend on the sequence homology between the antisense and the target genes. In the unevenly coloured inflorescences detected among anti-gchs1 transformants during their growth, relaxation of the antisense effect was here shown to start from the most distant member of the gene family, further demonstrating the influence of sequence homology in the stability of antisense inhibition.
Similar content being viewed by others
References
Bird CR, Ray JA, Fletcher JD, Boniwell JM, Bird AS, Teulieres C, Blan I, Bramley PM, Schuch W: Using antisense RNA to study gene function: inhibition of carotenoid biosynthesis intransgenic tomatoes. Bio/technology 9: 635–639 (1991).
Bourque JE: Antisense strategies for genetic manipulation in plants. Plant Sci 105: 125–149 (1995).
De Lange P, de Boer G-J, Mol JNM, Kooter JM: Conditional inhibition of β-glucuronidase expression by antisense gene fragments in petunia protoplasts. Plant Mol Biol 23: 45–55 (1993).
Elomaa P, Honkanen J, Puska R, Seppänen P, Helariutta Y, Mehto M, Kotilainen M, Nevalainen L, Teeri TH: Agrobacterium-mediated transfer of antisense chalcone synthase cDNA to Gerbera hybrida inhibits flower pigmentation. Bio/technology 11: 508–511 (1993).
Halpin C, Knight ME, Foxon GA, Campbell MM, Boudet AM, Boon JJ, Chabbert B, Tollier M-T, Schuch W: Manipulation of lignin quality by downregulation of cinnamyl alcohol dehydrogenase. Plant J 6: 339–350 (1994).
Hamilton AJ, Lycett GW, Grierson D: Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature 346: 284–286 (1990).
Helariutta Y, Elomaa P, Kotilainen M, Seppänen P, Teeri TH: Cloning of cDNA coding for dihydroflavonol-4-reductase (DFR) and characterization of dfr expression in the corollas of Gerbera hybrida var. Regina (Compositae). Plant Mol Biol 22: 183–193 (1993).
Helariutta Y, Elomaa P, Kotilainen M, Griesbach RJ, Schröder J, Teeri TH: Chalcone synthase like genes active during corolla development are differentially expressed and encode enzymes with different catalytic properties in Gerbera hybrida (Compositae). Plant Mol Biol, in press (1995).
Kohno-Murase J, Murase M, Ichikawa H, Imamura J: Effects of an antisense napin gene on seed storage compounds in transgenic Brassica napus seeds. Plant Mol Biol 26: 1115–1124 (1994).
Mol JNM, van der Krol AJ, van Tunen R, van Blokland R, de Lange P, Stuitje AR: Regulation of plant gene expression by antisense RNA. FEBS Lett 268: 427–430 (1990).
Neuhaus J-M, Flores S, Keefe D, Ahl-Goy P, Meins FJr: The function of vacuolar β-1,3-glucanase investigated by antisense transformation. Susceptibility of transgenic Nicotiana sylvestris plants to Cercospora nicotianae infection. Plant Mol Biol 19: 803–813 (1992).
Oeller PW, Lu M-W, Taylor LP, Pike DA, Theologis A: Reversible inhibition of tomato fruit senescence by antisense RNA. Science 254: 437–439 (1991).
Peñarrubia L, Aguilar M, Margossian L, Fischer RL: An antisense gene stimulates ethylene hormone production during tomato fruit ripening. Plant Cell 4: 681–687 (1992).
Platt SG, Yang N-S: Dot assay for neomycin phosphotransferase activity in crude cell extracts. Anal Biochem 162: 529–535 (1987).
Rodermel SR, Abbott MS, Bogorad L: Nuclear-organelle interactions: nuclear antisense gene inhibits ribulose biphosphate carboxylase enzyme levels in transformed tobacco plants. Cell 55: 673–681 (1988).
Salehuzzaman SNIM, Jacobsen E, Visser RGF: Isolation and characterization of a cDNA encoding granulebound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potato. Plant Mol Biol 23: 947–962 (1993).
Sheehy RE, Kramer M, Hiatt WR: Reduction of polygalacturonase activity in tomato fruit by antisense RNA. Proc Natl Acad Sci USA 85: 8805–8809 (1988).
Smith CJS, Watson CF, Ray J, Bird CR, Morris PC, Schuch W, Grierson D: Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes. Nature 334: 724–726 (1988).
Smith CJS, Watson CF, Morris PC, Bird CR, Seymour GB, Gray JE, Arnold C, Tucker GA, Schuch W, Harding S, Grierson D: Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes. Plant Mol Biol 14: 369–379 (1990).
Tieman DM, Harriman RW, Ramamohan G, Handa AK: An antisense pectin methylesterase gene alters pectin chemistry and soluble solids in tomato fruit. Plant Cell 4: 667–679 (1992).
van Blokland R, van der Geest N, Mol JNM, Kooter JM: Transgene-mediated suppression of chalcone synthase expression in Petunia hybrida results from an increase in RNA turnover. Plant J 6: 861–877 (1994).
van der Krol AR, Lenting PE, Veenstra J, van der Meer IM, Koes RE, Gerats AGM, Mol JNM, Stuitje AR: An anti-sense chalcone synthase gene in transgenic plants inhibits flower pigmentation. Nature 333: 866–869 (1988).
van der Krol AR, Mur LA, de Lange P, Gerats AGM, Mol JNM, Stuitje AR: Antisense chalcone synthase genes in petunia: visualization of variable transgene expression. Mol Gen Genet 220: 204–212 (1990).
van der Meer IM, Stam ME, van Tunen AJ, Mol JNM, Stuitje AR: Antigense inhibition of flavonoid biosynthesis in petunia anthers results in male sterility. Plant Cell 4: 253–262 (1992).
Vaucheret H, Kronenberger J, Lepingle A, Vilaine F, Boutin J-P, Caboche M: Inhibition of tobacco nitrite reductase activity by expression of antisense RNA. Plant J 2: 559–569 (1992).
Visser RGF, Somhorst I, Kuipers GJ, Ruys NJ, Feenstra WJ, Jacobsen E: Inhibition of the expression of the gene for granule-bound starch synthase in potato by antisense constructs. Mol Gen Genet 225: 289–296 (1991).
Zhang H, Scheirer DC, Fowle WH, Goodman HM: Expression of antisense or sense RNA of an ankyrin repeatcontaining gene blocks chloroplast differentiation in Arabidopsis. Plant Cell 4: 1575–1588 (1992).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Elomaa, P., Helariutta, Y., Kotilainen, M. et al. Transformation of antisense constructs of the chalcone synthase gene superfamily into Gerbera hybrida: differential effect on the expression of family members. Mol Breeding 2, 41–50 (1996). https://doi.org/10.1007/BF00171350
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00171350