Hostname: page-component-586b7cd67f-gb8f7 Total loading time: 0 Render date: 2024-11-27T14:28:10.021Z Has data issue: false hasContentIssue false
  • English
  • Français

Osmotin-regulated reserve accumulation and germination in genetically transformed tea somatic embryos: a step towards regulation of stress tolerance and seed recalcitrance1

Published online by Cambridge University Press:  22 February 2007

Amita Bhattacharya ,
Uksha Saini ,
Preeti Sharma ,
P.K. Nagar  and
Paramvir Singh Ahuja
Amita Bhattacharya*
Affiliation:
Division of Biotechnology, Institute of Himalayan Bioresource Technology, Palampur, -176061, Himachal Pradesh, India
Uksha Saini
Affiliation:
Division of Biotechnology, Institute of Himalayan Bioresource Technology, Palampur, -176061, Himachal Pradesh, India
Preeti Sharma
Affiliation:
Division of Biotechnology, Institute of Himalayan Bioresource Technology, Palampur, -176061, Himachal Pradesh, India
P.K. Nagar
Affiliation:
Division of Biotechnology, Institute of Himalayan Bioresource Technology, Palampur, -176061, Himachal Pradesh, India
Paramvir Singh Ahuja
Affiliation:
Division of Biotechnology, Institute of Himalayan Bioresource Technology, Palampur, -176061, Himachal Pradesh, India
*
*Correspondence: Email: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

The osmotin gene was introduced into tea [Camellia sinensis (L.) O. Kuntze] somatic embryos at the globular stage of development via microprojectile bombardment, to determine the effects of this stress-tolerance-related gene on the accumulation of storage reserves and acquisition of desiccation tolerance during embryo maturation. Changes in total soluble sugars, starch and proteins in response to osmotin were estimated in the stably transformed embryos, using standard biochemical and histochemical methods, after the globular embryos had matured to the heart stage of development. These changes also were compared with two types of controls, comprising (1) untransformed embryos and (2) embryos that were bombarded without osmotin DNA. The control embryos showed poor reserve accumulation and negligible germination but, in contrast, the embryos bombarded with the osmotin gene showed a several-fold increase in storage reserves, normal maturation and 57–63% germination. When these transformed embryos were desiccated to 15.5% moisture content, only 40% germinated, although a further increase in the accumulation of storage reserves occurred. In contrast, the control embryos became necrotic and failed to germinate. This study demonstrates the use of somatic embryogenesis as a tool for understanding seed recalcitrance in tea, and suggests that transformation with stress-tolerance genes can overcome the developmental problems associated with recalcitrant seeds.

Keywords

Camellia sinensisdesiccation tolerancegerminationmaturationosmotinrecalcitrant seedsreserve accumulationsomatic embryogenesistea
Type
Research Article
Information
Seed Science Research , Volume 16 , Issue 3 , September 2006 , pp. 203 - 211
Copyright
Copyright © Cambridge University Press 2006

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Adams, C.A., Rinnie, R.W. and Fjerstad, M.C. (1980) Starch deposition and carbohydrate activities in developing and germinating soya bean seeds. Annals of Botany 45, 577–582.CrossRefGoogle Scholar
Bhattacharya, A. and Ahuja, P.S. (2003) Prospects of transgenics in tea crop improvement. pp. 115–130. in Singh, R.P.;, Jaiwal, P.K. (Eds) Plant genetic engineering: Improvement of commercial plants, Vol. 3. Houston, Texas Sci Tech Publishing.Google Scholar
Bhattacharya, A., Nagar, P.K. and Ahuja, P.S. (2002) Seed development in Camellia sinensis (L.) O. Kuntze. Seed Science Research 12, 39–46.CrossRefGoogle Scholar
Bradford, M.M. (1976) A rapid method for quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Analytical Biochemistry 72, 248–254.CrossRefGoogle ScholarPubMed
Doyle, J.J. and Doyle, J.L. (1990) Isolation of DNA from fresh plant tissue. Focus 12, 13–15.Google Scholar
Grover, A., Kapoor, A., Lakshmi, O.S., Agarwal, S., Sahi, C., Katiyar-Agarwal, S., Agarwal, M. and Dubey, H. (2001) Understanding molecular alphabets of the plant abiotic stress responses. Current Science 80, 206–216.Google Scholar
Ingram, J. and Bartels, D. (1996) The molecular basis of dehydration tolerance in plants. Annual Review of Plant Physiology and Plant Molecular Biology 47, 377–403.CrossRefGoogle ScholarPubMed
ISTA (International Seed Testing Association) (1985) Determination of moisture content. Seed Science and Technology 13, 338–341.Google Scholar
Jankun, J., Selman, S.H., Swiercz, R., Skrzypczak-Jankun, E. (1997) Why drinking green tea could prevent cancer. Nature 387, 561CrossRefGoogle ScholarPubMed
Kermode, A.R., Bewley, J.D., Dasgupta, J. and Misra, S. (1986) The transition from seed development to germination: a key role for desiccation?. HortScience 21, 1113–1118.CrossRefGoogle Scholar
McCready, R.M., Guggolz, J., Silviera, V. and Owens, H.S. (1950) Determination of starch and amylose in vegetables: Application to peas. Analytical Chemistry 22, 1156–1158.CrossRefGoogle Scholar
McKersie, B.D., Senaratna, T. and Bowley, S.R. (1990) Drying somatic embryos for use as artificial seeds. Proceedings of the Plant Growth Regulator Society of America 17, 199–207.Google Scholar
Mondal, T.K. (1999) Studies on RAPD markers for detection of genetic diversity, in vitro regeneration and Agrobacterium mediated genetic transformation of tea (Camellia sinensis L. (O.) Kuntze. PhD thesis, submitted to Utkal University, Orissa, India.Google Scholar
Mondal, T.K., Bhattacharya, A., Sood, A. and Ahuja, P.S. (2000) Factors affecting induction and cold storage of encapsulated tea Camellia sinensis (L.) O. Kuntze somatic embryos. Tea 21, 92–100.Google Scholar
Mondal, T.K., Bhattacharya, A. and Ahuja, P.S. (2001) Induction of synchronous secondary embryogenesis in Camellia sinensis (L.) O. Kuntze. Journal of Plant Physiology 158, 945–951.CrossRefGoogle Scholar
Mondal, T.K., Bhattacharya, A., Sood, A. and Ahuja, P.S. (2002) Factors affecting germination and conversion frequency of somatic embryos of tea Camellia sinensis (L.) O. Kuntze somatic embryos. Journal of Plant Physiology 159, 1317–1321.CrossRefGoogle Scholar
Pla, M., Huguet, G., Verdaguer, D., Puigderrajols, P., Llompart, B., Nadal, A. and Molinas, M. (1998) Stress proteins co-expressed in suberized and lignified cells and in apical meristems. Plant Science 139, 49–57.CrossRefGoogle Scholar
Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular cloning – A laboratory manual 2nd edition. Cold Spring Harbor, New York, Cold Spring Harbor Laboratory Press.Google Scholar
Sandal, I., Bhattacharya, A. and Ahuja, P.S. (2004) Transgenic tea through biolistic using leaf explants. US patent application 20040216191, 28 October 2004.Google Scholar
Sharma, P. (2003) Desiccation tolerance studies in tea somatic embryos. MSc Thesis, Chaudhary Sarwan Kumar Himachal Pradesh Krishi Vishvavidyalaya, PalampurGoogle Scholar
Sharma, P., Pandey, S., Bhattacharya, A., Nagar, P.K. and Ahuja, P.S. (2004) ABA associated biochemical changes during somatic embryo development in Camellia sinensis (L.) O. Kuntze. Journal of Plant Physiology 161, 1269–1276.CrossRefGoogle ScholarPubMed
Singh, N.K., Bracker, C.A., Hasegawa, P.M., Handa, A.K., Buckel, S., Hermodson, M.A., Pfankoch, E., Regnier, F.E. and Bressan, R.A. (1987) Characterization of osmotin. A thaumatin-like protein associated with osmotic adaptation in plant cells. Plant Physiology 85, 529–536.CrossRefGoogle ScholarPubMed
Sood, A., Palni, L.M.S., Sharma, M., Rao, D.V., Chand, G. and Jain, N.K. (1993) Micropropagation of tea using cotyledon culture and encapsulated somatic embryos. Journal of Plantation Crops 21, 295–300.Google Scholar
Töpfer, R., Schell, J., Steinbiss, H.-H. (1988) Versatile cloning vectors for transient gene expression and direct gene transfer in plant cells. Nucleic Acids Research 16, 8725CrossRefGoogle ScholarPubMed
Vieitez, A.M. (1995) Somatic embryogenesis in Camellia spp. pp. 235–276. in Jain, S.M.;, Gupta, P.K.;, Newton, R. (Eds) Somatic embryogenesis in woody plants, Vol. 2, Angiosperms. Dordrecht, Kluwer Academic Publishers.CrossRefGoogle Scholar