Skip to main content Accessibility help

Evaluating biological containment strategies for pollen-mediated gene flow

  • Alexandra Hüsken, Sabine Prescher and Joachim Schiemann


Several biological containment methods have been developed to reduce pollen dispersal; many of them only have a proof of concept in a model plant species. This review focuses on biological containment measures which were tested for their long-term efficiency at the greenhouse or field scale level, i.e. plastid transformation, transgene excission, cleistogamy and cytoplasmic male sterility (CMS). Pollen-mediated gene transfer in transplastomic tobacco could occur at very low frequencies if the predominant mode of inheritance is maternal. Transgene excision from tobacco pollen can be made highly efficient by coexpression of two recombinases. For cleistogamous oilseed rape it was shown that some flowers were partially open depending on genotypes, environment and recording dates. Reports on the stability of CMS in maize and sunflower indicated that there is a high variability for different genotypes under different environmental conditions and over successive years. But for both crop types some stable lines could be selected. These data demonstrate that the biological containment methods discussed are very promising for reducing gene flow but that no single containment strategy provides 100% reduction. However, the necessary efficiency of biological containment methods depends on the level of containment required. The containment level may need to be higher for safety purposes (e.g. production of special plant-made pharmaceuticals), while much lower containment levels may already be sufficient to reach coexistence goals. It is concluded that where pollen-mediated gene flow must be prevented altogether, combinations of complementary containment systems will be required.

    • Send article to Kindle

      To send this article to your Kindle, first ensure is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about sending to your Kindle. Find out more about sending to your Kindle.

      Note you can select to send to either the or variations. ‘’ emails are free but can only be sent to your device when it is connected to wi-fi. ‘’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

      Find out more about the Kindle Personal Document Service.

      Evaluating biological containment strategies for pollen-mediated gene flow
      Available formats

      Send article to Dropbox

      To send this article to your Dropbox account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Dropbox.

      Evaluating biological containment strategies for pollen-mediated gene flow
      Available formats

      Send article to Google Drive

      To send this article to your Google Drive account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Google Drive.

      Evaluating biological containment strategies for pollen-mediated gene flow
      Available formats


Corresponding author

*Corresponding author:


Hide All
[1]Allainguillaume, J, Harwood, T, Ford, CS, Cuccato, G, Norris, C, Allender, CJ, Welters, R, King, GJ, Wilkinson, MJ (2009) Rapeseed cytoplasm gives advantage in wild relatives and complicates genetically modified crop biocontainment. New Phytol. 183: 12011211
[2]Avni, A, Edelmann, M (1991) Direct selection for paternal inheritance of chloroplasts in sexual progeny of Nicotiana. Mol. Gen. Genet. 225: 273277
[3]Azhagiri, AK, Maliga, P (2007) Exceptional paternal inheritance of plastids in Arabidopsis suggests that low-frequency leakage of plastids via pollen may be universal in plants. Plant J. 52: 817823
[4]Benitez, ER, Khan, NA, Matsumura, H, Abe, J, Takahas, R (2010) Varietal differences and morphology of cleistogamy in soybean. Crop Sci. 50: 185190
[5]Birky, CW (1995) Uniparental inheritance of mitochondrial and chloroplast genes: Mechanisms and evolution. Proc. Natl. Acad. Sci. USA 92: 1133111338
[6]Bock, R, Khan, MS (2004) Taming plastids for a green future. Trends Biotechnol. 22: 311318
[7]Bock, R, Timmis, JN (2008) Reconstructing evolution: gene transfer from plastids to the nucleus. Bioassays 30: 556566
[8]Budar, F, Touzet, P, De Paepe, R (2003) The nucleo-mitochondrial conflict in cytoplasmic male sterilities revisited. Genetica 117: 316
[9]Chase, CD (2006) Genetically engineered cytoplasmic male sterility. Trends Plant Sci. 11: 79
[10]Chase, CD (2007) Cytoplasmic male sterility: a window to the world of plantmitochon-drial-nuclear interaction. Trends Genet. 23: 8190
[11]Chase CD, Gabay-Laughnan S (2004) Cytoplasmic male sterility and fertility restoration by nuclear genes. In Daniell H, Chase CD, eds, Molecular Biology and Biotechnology of Plant Organelles, Springer, New York, pp 593–622
[12] Cilier, M, Feruzan, D, Göksel, O (2004) Histological aspects of anther wall in male fertile and cytoplasmic male sterile Helianthus annuus L. Asi. J. Plant Sci. 3: 145150
[13]Connor, HE (1979) Breeding systems in the grasses: a survey. New Zealand Journal of Botany 17: 547574
[14] Conner, AJ, Glare, TR, Nap, JP (2003) The release of genetically modified crops into the environment – Part II. Overview of ecological risk assessment. Plant J. 33: 1946
[15] Cummins, JE (1998) Chloroplast-transgenic plants are not a gene flow panacea. Nat. Biotechnol. 16: 401
[16]Daniell, H (2002) Molecular strategies for gene containment in transgenic crops. Nat. Biotechnol. 20: 581586
[17]Daniell, H, Kumar, S, Dufourmantel, N (2005) Breakthrough in chloroplast genetic engineering of agronomically important crops. Trends Biotechnol. 23: 238245
[18]De Maagd RA, Boutilier K (2009) Efficacy of strategies for biological containment of transgenic crops. A literature review. Plant Research International, Note 650
[19]DeCosa, B, Moar, W, Lee, SB, Miller, M, Daniell, H (2001) Overexpression of the Bt cry2Aa2 operon in chloroplasts leads to formation of insecticidal crystals. Nat. Biotechnol. 19: 7174
[20]Den Nijs HCM, Bartsch D, Sweet J (eds) (2004) Introgression from genetically modified plants into wild relatives. CABI, p 432
[21]Derepas, A, Dulieu, H (1992) Inheritance of the capacity to transfer plastids by the pollen parent in Petunia hybrida hort. Heredity 83: 610
[22] Diaz, A, MacNair, MR (1998) The effect of plant size on the expression of cleistogamy in Mimulus nasutus. Funct. Ecol. 12: 92–98
[23]Dong, J, Wagner, DB (1994) Paternally inherited chloroplast polymorphism in Pinus: estimation of diversity and population subdivision, and tests of disequilibrium with maternally inherited mitochondrial polymorphism. Genetics 136: 11871194
[24]Dunwell JC, Ford CS (2005) Technologies for biological containment of GM- and non-GM-crops. Final Report, Defra Contract CBEC 47
[25]EFSA (2009) Scientific opinion on guidance for the risk assessment of genetically modified plants used for non-food or non-feed purposes. EFSA Journal 1164: 1–42
[26] Fan, ZG, Stefansson, BR (1986) Influence of temperature on sterility of 2 cytoplasmic male-sterility systems in rape (Brassica napus L.). Can J. Plant Sci. 66: 221227
[27]Gealy DR (2005) Gene movement between rice (Oryza sativa) and weedy rice (Oryza sativa): a U.S. temperate rice perspective. In Gressel J., ed, Crop Ferality and Volunteerism, CRC Press
[28]Gidoni, D, Srivastava, V, Carmi, N (2008) Site-specific excisional recombination strategies for elimination of undesirable transgenes from crop plants. In Vitro Cell Dev. Biol. - Plant 44: 457467
[29] Gressel, J, Al-Ahmad, H (2005) Assessing and managing biological risks of plants used for bioremediation, including risks of transgene flow. Z. Nat. forsch. 60c: 154165
[30]Gressel, J, Valverde, BE (2009) A strategy to provide long-term control of weedy rice while mitigating herbicide resistance transgene flow, and its potential use for other crops with related weeds. Pest. Manag. Sci. 65: 723731
[31]Grevich, JJ, Daniell, H (2005) Chloroplast genetic engineering: Recent advances and future perspectives. Crit. Rev. Plant Sci. 24: 83107
[32]Havey M (2004) The use of cytoplasmic male-sterility in hybrid seed production. In Daniell H, Chase C, eds, Molecular Biology and Biotechnology of Plant Organelles, Springer, New York, pp 623–634
[33] Haygood, RA, Ives, AR, Andow, DA (2004) Population genetics of transgene containment. Ecol. Lett. 7: 213220
[34]Hoshikawa K (1993) Anthesis, fertilization and development of caryopsis. In Matsuo T, Hoshikawa K, eds, Science of the rice plant, vol. 1 (Morphology), Food and Agriculture Policy Research Center, Nobunkyo Publ. Co, Tokyo, pp 339–376
[35]Hüsken, A, Dietz-Pfeilstetter, A (2007) Pollen-mediated gene flow from herbicide-resistant oilseed rape (Brassica napus L.). Trans. Res. 16: 557569
[36]Hvarleva, T, Hristova, M, Bakalova, A, Hristov, M, Atanossov, I, Atanassov, A (2009) CMS lines for evaluation of pollen flow in sunflower relevance for transgene flow mitigation. Biotechnol. Biotechnol. Equip. 23: 13091315
[37]Kriete, G, Niehaus, K, Perlick, AM, Puhler, A, Broer, I (1996) Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compound N-acetyl-L-phosphinothricin. Plant J. 9: 809818
[38] Kuraichi, N, Makino, T, Hirose, S (1994) Inheritance of cleistogamy-chasmogamy in barley. Barley Genet. Newsl. 23: 19
[39]Latha, R, Thiyagarajan, K, Senthilvel, S (2004) Genetics, fertility behavior and molecular marker analysis of a new TGMS line, TS6, in rice. Plant Breed. 123: 235240
[40]Leflon M, Pinochet X, Hüsken A, Pendergast D, Knightly S (2009a) Cleistogamy of oilseed rape: a way to prevent pollen flow at the field scale ? In 5th ISHS International Symposium on Brassicas, Programme and Abstract Book, Lillehammer, Norway
[41]Leflon, M, Hüsken, A, Njontje, C, Knightly, S, Pendergast, D, Pierre, J, Renard, M, Pinochet, X (2009b) Stability of the cleistogamous trait during the flowering period of oilseed rape. Plant Breed. 129: 1318
[42]Lu, BR (2003) Transgene containment by molecular means – is it possible and cost effective? Environ. Biosafety Res. 2: 38
[43]Lu, BR (2008) Transgene escape from GM crops and potential biosafety consequences: An anvironmental perspective. Coll. Biosafety Reviews 4: 66141
[44]Lu YH, Belcram H, Rouault P, Piel N, Lucas MO, Falentin C, Renard M, Chalhoub B, Delourme R (2008) Cloning of a cleistogamy gene Clg1 in oilseed rape (B. napus L). In 5th ISHS International Symposium on Brassicas, Programme and Abstract Book, Lillehammer, Norway
[45] Luo, K, Duan, H, Zhao, D, Zheng, X, Deng, W, Chen, Y, Stewart, Jr CN, McAvoy, R, Jiang, X, Wu, Y, He, A, Pei, Y, Li, Y (2007) ‘GM-gene-deletor’: Fused loxP-FRT recognition sequences dramatically improve the efficiency of FLP or CRE recombinase on transgene excision from pollen and seed of tobacco plants. Plant Biotechnol. J. 5: 263274
[46]Marshall, DR, Thomson, NJ, Nichols, GH, Patrick, CM (1974) Effects of temperature and day length on cytoplasmic male sterility in Cotton (Gossypium). Aust. J. Agric. Res. 25: 443450
[47]Medgyesy, P, Menczel, L, Maliga, P (1980) The use of cytoplasmic streptomycin resistance: chloroplast transfer from Nicotiana tabacum into Nicotiana sylvestris, and isolation of their somatic hybrids. Mol. Gen. Genet. 179: 693698
[48]Medgyesy, P, Páy, A, Márton, L (1986) Transmission of paternal chloroplasts in Nicotiana. Mol. Gen. Genet. 204: 195198
[49]Miyashita, K, Matsuda, H, Ohara, M, Misawa, T, Shimamoto, Y (1999) Flowering and fruiting dynamics of chasmogamous and cleistogamous flowers in wild and cultivated soybeans. Res. Bul. Univ. Farm 31: 4148
[50]Mlynárová, L, Conner, A, Nap, JP (2006) Directed microspore-specific recombination of transgenic alleles to prevent pollen-mediated transmission of transgenes. Plant Biotechnol. J. 4: 445452
[51]Munsch, M, Camp, KH, Stamp, P, Weider, C (2008) Modern maize hybrids can improve grain yield as plus-hybrids by the combined effects of cytoplasmic male sterility and allo-pollination. Maydica 53: 262268
[52] Munsch, M, Stamp, P, Christov, NK, Foueillassar, XM, Hüsken, A, Camp, KH, Weider, C (2010) Grain yield increase and pollen containment by Plus-Hybrids could improve acceptance of transgenic maize. Crop Sci. 50: 909919
[53]Murphy, DJ (2007) Improving containment strategies in biopharming. Plant Biotechnol J. 5: 555569
[54]Nagao, S, Takahashi, M (1963) Trial construction of twelve linkage groups in Japanese rice. Genetical studies on rice plants. J. Fac. Agr. Hokkaido Univ. 53: 72130
[55] Pelletier, G, Budar, F (2007) The molecular biology of cytoplasmically inherited male sterility and prospects for its engineering. Curr. Opin. Biotechnol. 18: 121125
[56] Perez-Prat, E, van Lockeren Campagne, MM (2002) Hybrid seed production and the challenge of propagating male-sterile plants. Trends Plant Sci. 7: 199203
[57]Peterson, CE, Foskett, RL (1953) Occurrence of pollen sterility in seed fields of Scott County Globe onions. Proc. Am. Soc. Hort. Sci. 62: 443448
[58] Peterson, G, Cunningham, S, Deutsch, L, Erikson, J, Quinlan, A, Raez-Luna, E, Tinch, R, Troell, M, Woodburg, P, Zens, S (2000) The risks and benefits of genetically modified crops: a multidisciplinary perspective. Conserv. Ecol. 4: 13
[59]Quesada-Vargas, T, Ruiz, ON, Daniell, H (2005) Characterization of heterologous multigene operons in transgenic chloroplasts. Plant Physiol. 138: 17461762
[60]Ruf, S, Karcher, D, Bock, R (2007) Determining the transgene containment level provided by chloroplast transformation. Proc. Natl. Acad. Sci. USA 104: 69987002
[61] Ruiz, O, Daniell, H (2005) Cytoplasmic male sterility engineered via the plastid genome. Plant Physiol. 138: 12321246
[62]Sarvella, P (1966) Environmental influences on sterility in cytoplasmic male-sterile cottons. Crop Sci. 6: 361364
[63]Sawhney, VK (2004) Photoperiod-sensitive male sterile mutant in tomato and its potential use in hybrid seed production. J. Hortic. Sci. Biotech. 79: 138141
[64] Sheppard, AE, Ayliffe, MA, Blatch, L, Day, A, Delaney, SK, Khairul-Fahmy, N, Li, Y, Madesis, P, Pryor, AJ, Timmis, JN (2008) Transfer of plastid DNA to the nucleus is elevated during male gametogenesis in tobacco. Plant Phys. 148: 328336
[65]Stamp, P, Chowchong, S, Menzi, M, Weingartner, U, Kaeser, O (2000) Increase in the yield of cytoplasmic male sterile maize revisited. Crop Sci. 40: 15861587
[66]Stegemann, S, Bock, R (2006) Experimental reconstruction of functional gene transfer from the tobacco plastid genome to the nucleus. Plant Cell 18: 28692878
[67]Svab, Z, Maliga, P (2007) Exceptional transmission of plastids and mitochondria from the transplastomic pollen parent and its impact on transgene containment. Proc. Natl. Acad. Sci. USA 104: 70037008
[68]Testolin, R, Cipriani, G (1997) Paternal inheritance of chloroplast DNA and maternal inheritance of mitochondrial DNA in the genus Actinidia. Theor. Appl. Genet. 94: 897903
[69]Turuspekov, Y, Honda, I, Watanabe, Y, Stein, N, Komatsuda, T (2009) An inverted and micro-colinear genomic regions of rice and barley carrying the cly1 gene for cleistogamy. Breed. Science 59: 657663
[70] Wang, T, Li, Y, Shi, Y, Reboud, X, Darmency, H, Gressel, J (2004) Low frequency transmission of a plastid-encoded trait in Setaria italica. Theor. Appl. Genet. 108: 315320
[71]Weingartner, U, Kaeser, O, Long, M, Stamp, P (2002) Combining cytoplasmic male sterility and xenia increases grain yield of maize hybrids. Crop Sci. 42: 18481856
[72]Weider, C, Stamp, P, Christov, N, Hüsken, A, Foueillassar, X, Camp, KH, Munsch, M (2009) Stability of cytoplasmic male sterility in maize under different environmental conditions. Crop Sci. 49: 7784
[73]Yoshida, G, Itoh, JI, Ohmori, S, Miyoshi, K, Horigome, A, Uchida, E, Kimizu, M, Matsumura, Y, Kusaba, M, Satoh, H, Nagato, Y (2007) Superwoman1-cleistogamy, a hopeful allele for gene containment in GM-rice. Plant Biotechnol. J. 5: 112


Related content

Powered by UNSILO

Evaluating biological containment strategies for pollen-mediated gene flow

  • Alexandra Hüsken, Sabine Prescher and Joachim Schiemann


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.