Abstract
The seaweed Laminaria japonica (Phaeophyceae) has a two-generation life cycle consisting of haploid gametophytes and diploid sporophytes. Female and/or male gametophytes were transformed using particle bombardment and the histological LacZ assay was performed on sporophytes generated by either parthenogenesis or inbreeding. Female gametophyte-targeted transformation resulted in similar lower efficiencies in both parthenogenetic and zygotic sporophytes, and only a chimeric expression pattern was observed. Male gametophyte-targeted transformation led to a higher efficiency, with 3.5% of the zygotic sporophytes stained completely blue (all-blue), implying the integration of lacZ at the one-cell stage. Polymerase chain reaction analysis using primers specific for a lacZ-vector juncture fragment and subsequent blotting indicated the presence of the introduced gene in the sporophytes. The method reported here has a potential for seaweed transformation using spore-based bombardment followed by the developmental process.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- DPR :
-
Detected positive rate
- ER :
-
Expression rate
References
Alwen A, Eller N, Kastler M, Benito Maria MR, Heberle-Bors E (1990) Potential of in vitro pollen maturation for gene transfer. Physiol Plant 79:194–196
Dai JX, Fang ZX (1976) Some preliminary observations of the parthenogenesis in Laminaria japonica Aresch. (in Chinese with English abstract). Acta Genet Sin 3:32–38
Fang ZX (1983) A summary of the genetic studies of Laminaria japonica in China. In: Tseng CK (ed) Proc Joint China-US Phycol Symp. Science Press, Beijing, pp 123–136
Huang X, Weber JC, Hinson TK, Mathieson AC, Minocha SC (1996) Transient expression of the GUS reporter gene in the protoplasts and partially digested cells of Ulva lactuca L. (Chlorophyta). Bot Mar 39:467–474
Jiang P, Qin S, Tseng CK (2002) Expression of hepatitis B surface antigen gene (HBsAg) in Laminaria japonica (Laminariales, Phaeophyta). Chin Sci Bull 47:1438–1440
Kuang M, Wang SJ, Li Y, Shen DL, Tseng CK (1998) Transient expression of exogenous GUS gene in Porphyra yezoensis (Rhodophyta). Chin J Oceanol Limnol 16[Suppl]:56–61
Kübler JE, Minocha SC, Mathieson AC (1994) Transient expression of the GUS reporter gene in protoplast of Porphyra miniata (Rhodophyta). J Mar Biotechnol 1:165–169
Kurtzman AM, Cheney DP (1991) Direct gene transfer and transient expression in a marine red alga using the biolistic method. J Phycol 27[Suppl]:42
Maier I, Müller DG (1982) Antheridium fine structure and spermatozoid release in Laminaria digitata. Phycologia 21:1–8
Motomura T, Sakai Y (1984) Ultrastructural studies of gametogenesis in Laminaria angustata (Laminariales, Pheaophyta) regulated by iron concentration in the medium. Phycologia 23:331–343
Neuhaus G, Neuhaus-Url G, Gruss P, Schweiger H (1984) Enhancer-controlled expression of the simian virus 40 T-antigen in the green alga Acetabularia. EMBO J 3:2169–2172
Polne-Fuller M, Gibor A (1984) Developmental studies in Porphyra. I. Blade differentiation in Porphyra perforata as expressed by morphology, enzymatic digestion, and protoplast regeneration. J Phycol 20:609–616
Qin S, Zhang J, Li WB, Wang XH, Tong S, Sun YR, Tseng CK (1994) Transient expression of GUS gene in Phaeophytes using Biolistic Particle Delivery System (in Chinese with English abstract). Oceanol Liminol Sin 25:353–356
Qin S, Jiang P, Li XP, Tseng CK (1998a) The expression of lacZ in regenerated sporophytes of parthenogenetic Laminaria japonica. In: Menasveta P, Tanticharoen M (eds) Proc 2nd Asia-Pacific Marine Biotechnol Conf 3rd Asia-Pacific Conf Algal Biotechnol. BIOTEC Press, Bangkok, pp 205–208
Qin S, Jiang P, Li XP, Wang XH, Tseng CK (1998b) A transformation model for Laminaria japonica. Chin J Oceanol Limnol 16[Suppl]:50–55
Qin S, Sun GQ, Jiang P, Zou LH, Wu Y, Tseng CK (1999) Review of genetic engineering of Laminaria japonica (Laminariales, Phaeophyta) in China. Hydrobiologia 398/399:469–472
Reddy CRK, Migita S, Fujita Y (1989) Protoplast isolation and regeneration of three species of Ulva in axenic culture. Bot Mar 32:483–490
Renn D (1997) Biotechnology and the red seaweed polysaccharide industry: status, needs and prospects. Trends Biotechnol 15:9–14
Saunders GW (1993) Gel purification of red algal genomic DNA: an inexpensive and rapid method for the isolation of polymerase chain reaction-friendly DNA. J Phycol 29:251–254
Sawabe T, Ezura Y, Yamamoto H (1997) Plant regeneration from protoplasts of Laminaria japonica Areschoug (Laminariales, Phaeophyceae) in a continuous-flow culture system. Plant Cell Rep 17:109–112
Stevens DR, Purton S (1997) Genetic engineering of eukaryotic algae: progress and prospects. J Phycol 33:713–722
Acknowledgements
We would like to thank Prof. Fei Xiugeng of our institute for assistance in taking photographs.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by F. Sato
Rights and permissions
About this article
Cite this article
Jiang, P., Qin, S. & Tseng, C.K. Expression of the lacZ reporter gene in sporophytes of the seaweed Laminaria japonica (Phaeophyceae) by gametophyte-targeted transformation. Plant Cell Rep 21, 1211–1216 (2003). https://doi.org/10.1007/s00299-003-0645-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00299-003-0645-2