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Artificial Seeds for Phytoremediation

#K-1279.2


Development of Technology for Obtaining Artificial Seeds of Plants Heavy Metal-Hyperaccumulators for Phytoremediation of Polluted Soils

Tech Area / Field

  • ENV-RED/Remediation and Decontamination/Environment BIO-MIB/Microbiology/Biotechnology BIO-OTH/Other/Biotechnology

Status
3 Approved without Funding

Registration date
10.08.2007

Leading Institute
National Biotechnology Center of Kazakstan / Institute of Plant Biology and Biotechnology, Kazakstan, Almaty

Supporting institutes

  • Kazakh Scientific Center for Quarantine and Zoonotic Diseases, Kazakstan, Almaty

Collaborators

  • University of California / College of Agricultural and Environmental Sciences, USA, CA, Davis\nHumboldt-Universitat zu Berlin / Landwirtschaftlich-Gartnerische Fakultat, Germany, Berlin\nAlberta Agriculture, Food and Rural Development / Crop Diversification Centre South, Canada, Alberta\nAgriculture and Agri-Food Canada / Plant Gene resources of Canada, Saskatoon Research Centre, Canada, Saskatoon\nUniversity of Alberta / Department of Agricultural, Food and Nutrional Science, Canada, AB, Edmonton

Project summary

Full title of project “Development of technology for obtaining of the artificial seeds of heavy metal-hyperaccumulator plants for phytoremediation of polluted soils” ISTC.

Short title: Artificial Seeds for Phytoremediation

As a result of the ISTC К-487 project (2002-2004) «Cell selection of perspective plant species - heavy metal hyperaccumulators for phytoremediation of the environment from anthropogenic contamination” the cell lines with high tolerance to heavy metals (Cu ++, Cd ++) were obtained. Embryos and embryo-derived regenerated plants were obtained from the selected cells. Technical reports are attached. The next project “Development of technology for obtaining of the artificial seeds of heavy metal-hyperaccumulator plants for phytoremediation of polluted soils” is proposed for further 36 months.

Artificial (synthetic) seeds (AS) technology is the process of obtaining embryos without gamete fusion, somatic embryos. It provides large scale clonally propagation of new valuable, rare genotypes of plants.

AS are somatic embryos of the same stage of development that are placed in polymer capsule. The capsule protects embryos from unfavorable conditions, retains water, and provides sterility inside the capsule, which contains nutrition for initial growth of embryos. Capsule may be prepared from sodium alginate (McKersie and Brown, 1996).

AS technology offers many useful advantages on a commercial scale for the propagation of a variety of crop plants, especially crops for which true seeds are not used or expensive. This technology would also be useful for multiplying of the genetically engineered plants (transgenic plants), somatic and cytoplasmic hybrids, sterile and unstable genotypes. Besides, artificial seeds would be useful material for preservation of desirable elite genotypes (cryopreservation). They would also be valuable tools in experimental research to study the process of zygotic embryogenesis and understanding the role of endosperm in normal embryo development and germination.

The problem of antimicrobial properties of the capsule of AS is still unsolved (Lion, 1994).

Usage of AS in phytoremediation technology would greatly shorten the way (less than one year) from selected cell, embryos to exploring of modified genotypes in field; provide mass propagation of plant forms with useful properties. Besides this utilization of AS would allow to avoid recombination which occurres during pollination.

Generally the basic procedures of mass production and conservation of somatic embryos include the following stages:

  1. sterilization of explants and their cultivation on nutrient medium;
  2. dedifferentiation of cells and callus genesis;
  3. induction of somatic embryogenesis;
  4. sieving of the suspension and selection of proembryos;
  5. development of somatic embryos;
  6. drying of the embryos;
  7. incapsulation;
  8. germination and storing (McKersie at al., 1989).

This technology will allow mass, clonally propagation of cell lines derived in frame of the previous ISTC К-487 project independently from the season. Development of the AS production may begin from the already available cell line suspensions with high tolerance to HM.

Purpose of project: Laboratory and experimental/industrial protocol for obtaining of Medicago sativa artificial seeds from cells tolerant to heavy metals.

Expected Results and Their Application

This investigation relates to applied work. Laboratory and experimental/industrial protocol for obtaining of the AS-Medicago sativa from selected cells tolerant to heavy metals will be obtained. The AS of the HM hyperaccumulator plants would be used and purchased by many enterprises in Kazakhstan for phytoremediation of soils (Supporting letter is attached).

Competence of the project team in the specified area.

Institute of Plant Physiology, Genetics and Bioengineering. Laboratory of cell engineering. Head of the laboratory Dr. Uteulin K.R. Experience - cell selection, embryogenesis and regeneration. Publications:

Svanbaev E. Taukeleva Sh.N., Dzangalina E.D., Uteulin K.R. Embryogenic callus cultures. of alfalfa. Izvestia Academy Nauk of Kazakhstan. Biological series. 2002.N 1, P.65-69.

Uteulin K.R., Dzangalina E.D., Taukeleva Sh.N., Kalykova A.S., Sarsenbaev B.A. Influence of Cu++ ions on growth of suspension culture of alfalfa. VIII International Conference. The Biology of Plant Cells In Vitro and Biotechnology. Samara, Russia, September 9-13, 2003, P.338.

Laboratory of Root Nutrition. Head of the laboratory Professor Sarsenbaev B. Experience - control of heavy metal concentrations in plant organs and sub-cell fractions. Publications:

Kuzembaeva N.А., Sarsenbaev B.A. Release of cation -binding compounds from maize roots under osmotic shock. Plant Root Systems and Natural Vegetation. UPPSALA. 1996. P. 113-115 (English).

Sarsenbaev B.A., S.D. Atabaeva and B.A. Usenbekov B.A. Phytoremediation of waste water using free floating plants. Biotechnology. 2000, N1-2 (13). P. 103-106.

Kazak Scientific Center for Quarantine and Zoonotic Diseases (category 1). The head of control department is Professor Tugambayev T. Experience - methods of preventing infections of tissue cultures by viruses, bacteria’s and fungi.

Publications:

  1. Tugambaev T.I,. Matakov M. I,.Okulova. I.V - Oppression of anti complement properties of agar for reaction of immune hemolysis using polycationes. Biotechnology, theory and practice, 2002, N 4, P. 37-39.
  2. Tugambaev T.I., Dernovaya V.F. – Study of P. Multacida effect on antibody forming cells from some mice immunocempetence organs. News of AoSc, biology series, 2001, N 5, P. 81-85.
  3. Tugambaev T.I.,. Atshabar B.B,. Matakov M.I et al. – Manufacture of diagnostic immunoreagents for natural plague foci surveillance at the Republic of Kazakhstan. Astana medicinalyk zhurnaly, 2003, N 3, P. 116-119.

Meeting ISTC Goals and Objectives:
  • scientists from the Institute previously involved in weapon production would be oriented on peaceful activities;
  • integration with collaborator of the project;
  • supporting of biotechnologies in phytoremidation;
  • AS of the useful plant genotypes would be of high commercial value, the conversion to market economy corresponding to civil requirements would be achieved.

Role of Foreign Collaborators/Partners:
  • information exchange in the course of project implementation;
  • provide comments to the technical reports (quarterly, annual final, etc.), submitted by project participants to the ISTC;
  • testing and evaluation of equipment/technologies, developed in the course of the project;
  • conduction of joint seminars and workshops;
  • potential partnership in commercialization of results.

Technical Approach and Methodology

Object of investigation – The cell line (500) of Medicago sativa cv Krasnovodopadskaya, selected to 500 µM Cu++.The methods of tissue culture, growing of plants in laboratory and field conditions, differential centrifuging, atomic adsorption determination of heavy metals and microbiological methods will be used.


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