In Vitro Tissue Culture S tudies in Sunflower (Helianthus spp.)

In Vitro Tissue Culture S tudies in Sunflower (Helianthus spp.)

Sunflower (Helianthus annuus L.) is the fourth mos t important oilseed crop in the world in terms of total yearly production,after soybean, rapeseed, and groundnut. The classical breeding s tudies made from 1880 until today have beenfocused on major characters, such as high fat, pes ts and diseases resis tance. The narrow genetic base in sunflowercauses a major problem at breeding and selection of desired sunflower lines eg. cytoplasmic male s terility, disease andinsect pes t resis tance, fertility-res toration, agronomic and seed-oil characteris tics, drought tolerance, protein content,imidazolinone resis tance and fatty acid composition. The growth or maintenance of cells, tissues, organs, and theircomponents under defined physical and chemical conditions in vitro described as plant tissue culture, is an importanttool in both basic and applied researches as well as in commercial application. The firs t and important s tep of tissueculture is development of in vitro regeneration sys tem. Researchers have successful attempts by various explants, suchas immature embryo mature embryo, meris tem, anther, hypocotyl, protoplas t, cotyledonand embryo sac. Although thespecific techniques like, embryo culture, organogenesis, somatic embryogenesis have been succesfully used, antherculture, microspore culture ovule cultureneeds to be improved.

___

  • Abbas Mohamed S G El-D(2005). Genetic transformation of two high oleic Helianthus annuus L. genotypes using different transformation methods. Institute of Plant Molecular Physiology and Biotechnology, University of Bonn, Germany. PhD Thesis, pgs 179.
  • Alissa A, Serieys H and Jonard R(1985). Sur les possibilités de régénération d’espéces sauvages et d’hybrides interspécifiques du genre Helianthus par androgénèse in vitro. CRAcad. Sci. Paris 300, Série III, no. 1: 25–30.
  • Badea E, Prisecaru M and Angheluta H(1989). Studies on gynogenesis in intraespecific and interspecific hybrids in the genus Helianthus. Cercetari de Genetica Vegetala si Animala 1: l77-l83.
  • Berrios EF, Gentzbittel L, Serieys H, Alibert G and Sarrafi A(1999a). Influence of genotype and gelling agents on in vitro regeneration by organogenesis in sunflower. Plant Cell Tissue Org. Cult 59: 65-69.
  • Berrios EF, Gentzbittel L, Serieys H, Alibert G, Grievau Y, Berville A and Sarrafi A(1999b). Genetic control of in vitro organogenesis in recombinant inbred lines of sunflower (Helianthus annuus L). Plant Breed 118: 359-361.
  • Bhojwani SS, Razdan MK(1996). Chapter 11: Zygotic Embryo Culture. In: Plant Tissue Culture: Theory and Practice Volume 5, First Edition. Bhojwani SS, Razdan MK (eds.) Elsevier, Science B.V. The Netherlands. pp: 297-335.
  • Bohorova NE, Atanassov AI and Georgieva-Todorova J(1985). In vitro organogenesis, androgenesis and embryo culture in the Helianthus genus. Z. Pflanzenziicht 95: 35-44.
  • Bohorova NE, Cocking EC and Power JB(1986). Isolation, culture and callus regeneration of protoplasts of wild and cultivated Helianthusspecies. Plant Cell Rep 5: 256-258.
  • Breccia G, VegaT, Nestares G, Mayor ML, Zorzoli R, Picardi L(2009). Immature embryo culture for early screening of imidazolinone resistance in sunflower. Int. J of Plant Breed.3: 37-40.
  • Burrus M, Chanabe´C, Alibert G, BidneyD(1991). Regeneration of fertile plants from protoplasts of sunflower (Helianthus annuus L.). Plant Cell Rep 10:161-166.
  • Bürün B, Gürel A(2001). Embriyo kültürü. In: Bitki Biyoteknolojisi I, Doku Kültürü ve Uygulamaları, Babaoğlu M, Gürel E, Özcan S(eds),SÜ Basımevi, syf. 324-344.
  • Ceriani MF, Hopp HE, Hahne G and Escandon AS(1992). Cotyledons: An explant for routine regeneration of sunflower plants. Plant Cell Physiol 33(2): 157- 164.
  • Chanabé C, Burrus M, Bidney D and Alibert G(1991). Studies on plant regeneration from protoplasts in the genus Helianthus. Plant Cell Rep9: 635–638.
  • Chandler JM and Beard BH(1983). Embryo culture of Helianthus hybrids. Crop Sci 23:1004-1007.
  • Chandler JM and Jan CC(1985). Comparison of germination techniques for wild Helianthus seeds. Crop Sci 25: 356-358.
  • Chraibi KMB, Castelle JC, Latche A, Roustan J, Pand Fallot J(1992). A genotype independent system of regeneration from cotyledons of sunflower (Helianthus annuus L.). The role of ethylene. Plant. Sci 86: 215-221.
  • Christov M(2008). Helianthus species in breeding on sunflower. Proc. 17th Int. Sunfl. Conf., Cordoba, Spain, Int. Sunfl. Assoc., Paris, France 2: 709- 714.
  • Christov M(2012). Contribution of interspecific hybridization to sunflower breeding. Helia 35 (57): 37-46.
  • Christov M(2013). Contribution of interspecific and intergeneric hybridization to sunflower breeding. Helia 36(58): 1-18.
  • Cockerell TDA(1929) Hybrid sunflowers. Amer. Nat63: 470-475.
  • Coumans M and ZhongD(1995). Doubled haploid sunflower (Helianthus annuus) plant production by androgenesis: fact or artifact? Part 2. In vitro isolated microspore culture. Plant Cell, Tissue and Organ Culture 41: 203-209.
  • Dagustu N, Bayram G, Sincik M, Bayraktaroglu M(2012). The short breedıng cycle protocol effective on diverse genotypes of sunflower(Helianthus annuus L.). Turkish Journal of Field Crops17(2):124-128.
  • Dagustu N, Sincik M, Bayram G, Bayraktaroglu M(2010). Regeneration of fertile plants from sunflower (Helianthus annuus L.)-Immature embryo. Helia 33(52): 95-102.
  • Dağüstü N(2002). Some factors affecting the plant regeneration in sunflower (Helianthus annuus L.). Biotechnol and Biotechnol Eq 16(1): 18-23. Davey MR, Jan M(2010). Sunflower (Helianthus annuus L.): Genetic improvement using conventional and in vitro technologies. Journal of Crop Improvement 24 (4): 349-391.
  • Delgene L, Lesignes P, Alibert Gand Sarrafi A(1997). Genetic control of organogenesis in cotyledons of sunflower (Helianthus annuus). Plant Cell Tissue Org Cult 48: 127-130.
  • ElavazhaganT, Jayakumar S, Chitravadivu C and Balakrishnan V(2009). In vitro culture and cytological studies on Helianthus annus L. Botany Research International 2(4): 258–262.
  • Encheva J, Köhler H, Friedt W, Tsvetkova F, Ivanov P, Encheva V and Shindrova P(2003). Field evaluation of somaclonal variation in sunflower (Helianthus annuus L.) and its application for crop improvement. Euphytica 130: 167-175.
  • Encheva J, Tsvetkova F, Ivanov P(2004). Heritable tissue culture induced genetic variation in sunflower (Helianthus annuus L.) as a tool for crop improvement. Helia 27(41): 163-172.
  • Espinasse A and Lay C(1989). Shoot regeneration of callus derived from globular to torpedo embryos from 59 sunflower genotypes. Crop Sci 29: 201-205.
  • Faure N, Serieys H, Cazaux E, Kaan F, Bervillé A(2002). Partial hybridization in wide crosses between cultivated sunflower and the perennial Helianthus species H. mollis and H. orgyalis. Ann Bot 89: 31-39.
  • Fernandez-Martinez JM, Dominguez J, Perez-Vich B(2010). Update on breeding for resistance to sunflower broomrape. Helia 33(52): 1-12.
  • Finer JJ(1987). Direct somatic embryogenesis and plant regeneration from immature embryo of hybrid sunflower (Helianthus annuus L.) on high sucrose containing medium. Plant Cell Rep 6: 372-374.
  • Fiore MC, Trabace Tand Sunseri A(1997). High frequency of plant regeneration in sunflower from cotyledons via somatic embryogenesis. Plant Cell Reports 16: 295-298.
  • Fischer C, Klethi P and Hahne G(1992). Protoplasts from cotyledon and hypocotyl of sunflower (Helianthus annuus L.): Shoot Regeneration and Seed Production. Plant Cell Reports 11(12): 632–636 Flick CE, Evans DA and Sharp WR(1983). Organogenesis. In: Hand book of plant cell culture Vol. I. Techniques for propagation and breeding, EvansDA, SharpWR, Ammirato PVand Yamada Y(eds.) MacMillan New York, pp: 13-81.
  • Freyssinet M and Freyssinet G(1988). Fertile plant regeneration from sunflower (Helianthus annuus L) immature embryos. Plant. Sci 56: 177-181.
  • Gelebart P and San LH(1987). Obtention de plantes haploides par culture in vitro d’ovaires et d’ovules non fécondés de Tournesol (Helianthus annuus L.). Agronomie 7: 81–86.
  • Georgeiva-Todorova J, Bohorova N and Atanssov A(1980). Utilization of the method of tissue culture in the interspecific hybrids of Helianthus L Proc 9th Int Sunflower conference, Torremolinos, Spain, pp: 122-128.
  • Greco B, Tanzarella OA, Carrozzo Gand Blanco A(1984). Callus induction and shoot regeneration in sunflower. (Helianthus annuus L.). Plant Sci Lett 36: 73-77.
  • Guilley E and Hahne G(1989). Callus formation from isolated sunflower (H. annuus) mesophyll protoplasts. Plant Cell Reports 8: 226- 229.
  • Gürel A, Kontowski S, Nichterlein K and Friedt W(1991a). Embryogenesis in microspore culture of sunflower (Helianthus annuus L.). Helia 14: 123-128.
  • Gürel A, Nichterlein K and Friedt W(1991b). Shoot regeneration from anther culture of sunflower (Helianthus annuus L.) and some interspecific hybrids as affected by genotype and culture procedure. Plant Breeding 106: 68-76 Henderson JHM(1954). The changing nutritional pattern from normal to habituated sunflower callus tissue in vitro. Ann. Biol 30: 229-348.
  • Henn JJ, Wingender R and Schanbl H(1998). Regeneration of fertile, interspecific hybrids from protoplast fusions between Helianthus annuus L and wild Helianthus species. Plant Cell Rep 18: 220-224.
  • Henrickson CE(1954). The flowering of sunflower explants in aseptic culture. Plant Physiol 29: 536–538.
  • Hildebrandt AC, RikerAJ, Duggar BM(1946). The influence of the composition of the medium on growth in vitro of excised tobacco and sunflower tissue cultures. Am J Bot 33: 591-597.
  • Inoka K and Dahanayake N(2015). Effect of plant growth regulators on micro-propagation of sunflowe (Helianthus annuus L.). International Journal of Scientific and Research Publications 5(1): 1-5.
  • Jambhulkar SJ(1995). Rapid cycling through immature embryo culture in sunflower (Helianthus annuus L). Helia 18(22): 45-50.
  • Jan CC and Chandler JM(1985). Transfer of powdery mildew resistance from Helianthus debilis Nutt. to cultivated sunflower. Crop Science 25(4): 664- 666.
  • Jan CC(1988). Chromosome doubling of wild x cultivated sunflower interspecific hybrids and its direct effect on backcross success. In: Proc. Int. Sunflower Conf, 12th Novi Sad Yugoslavia. Int Sunf Assoc, Paris France, pp: 287–292.
  • Jeannin G and Hahne G(1991). Donor plant growth conditions and regeneration of fertile plants from somatic embryos induced on immature zygotic embryos. Plant Breed 107: 280-287.
  • Jeannin G, Bronner Rand Hahne G(1995) Somatic embryogenesis and organogenesis induced on the immature zygotic embryo of sunflower (Helianthus annuus L) cultivated in vitro: role of the sugar. Plant Cell Rep 15: 200-204.
  • Jonard R and Mezzarobba A(1990). Sunflower (Helianthus spp.): anther culture and field studies on haploids. In: Biotechnology in Agriculture and Forestry 10, Legumes and oilseed crops I, Bajaj YPS (ed.) Springer Verlag, Berlin, Heidelberg, New York Tokyo, pp: 485-501.
  • Kandler O(1952). Uber eine Physiologische Umstimmung Von Sonnenblumesntengelgewebe durch Dauereinwirkung Von-lndolylessigsaure. Planta 40: 346-349.
  • Karp A 1990. On the current understanding of somaclonal variation. Oxford Surveys of Plant Molecular and Cell Biology7: 1-58. Kräuter R, Steinmetz A and Friedt W(1991). Efficient interspecific hybridization in the genus Helianthus via ‘embro rescue’ and characterization of the hybrids. Theor Appl Genet 82: 521.
  • Kaya Y(2015). Herbicide resistance breeding in sunflower, current situation and future directions. Journal of ASM. Life Sciences. Plant Genetics and Breeding No. 2(326): 101-106.
  • Larkin PJ and Scowcroft WR(1981).Somaclonal variation-a novel source of variability from cell cultures for plant improvement. Theoretical and Applied Genetics60: 197-214. Lenee P and Chupeau Y(1986). Isolation and culture of sunflower protoplasts (Helianthus annuus L.): Factors influencing the viability of cell colonies derived from protoplasts. Plant Science43(1): 69- 75.
  • Lupi MC, Bennici A, Locciarid F, Gennai D(1987). Plantlet formation from callus and shoot tip culture of Helianthus annuus (L.). Plant Cell Tissue Org Cult 11: 47-55.
  • Malone-Schoneberg J, Scelonge CJ, Burrus M, and Bidney DL(1994). Stable transformation of sunflower using Agrobacterium and split embryogenic axis explants. Plant Sci 103: 199– 207.
  • Mandel JR, Dechaine JM, Marek LF, Burke JM(2011). Genetic diversity and population structure in cultivated sunflower and a comparison to its wild progenitor, Helianthus annuus L. Theor Appl Genet 123(5): 693-704.
  • Mezzarobba A, JonardR(1986). Effets du stade de prelevement et des pretraitements sur le developpement in vitro d’antheres preleves sur le tournesol cultive (Helianthus annus L.). CR Acad Sci Paris, 303: 181-186.
  • Miller JF and Al-Khatib K(2004). Registration of two oilseed sunflower genetic stocks, Sures-1 and Sures-2, resistant to tribenuron herbicide. Crop Sci 44:1037- 1038.
  • Mix G(1985). Antheren-und Ovarienkultur von Sonnenblumen (Helianthus annuus L.). Landbauforschung Voelkenrode 156–153 :35. Moghaddasi MS(2011). Sunflower Tissue Culture. Advances in Environmental Biology, 5(4): 746- 755.
  • Mohmand AS and Quraishi A(1994). Tissue culture of sunflower. Pakistan Journal of Agricultural Research 15(1): 153–160. Müller A, Iser M and Hess D(2001). Stable transformation of sunflower (Helianthus annuus L .) using a non-meristematic regeneration protocol and green fluorescent protein as a vital marker. Transgenic Research 10(1987): 435–444.
  • Nenova N, CristovM, Ivanov P(2000). Anther culture regeneration from some wild Helianthus species. Helia 23(32): 65-72. Nenova N, Valkova D, Encheva J, and Tahsin N(2014). Promising lines as a result from interspecific hybridization between cultivated sunflower (Helianthus annuus L.) and the perennial species Helianthus ciliaris. Turkish Journal of Agricultural and Natural SciencesSpecial Issue 2: 1654–1659.
  • Nestares G, Zorzoli R, Mroginski L and Picardi L(1996). Plant regeneration from cotyledons derived from mature sunflower seeds. Helia 19: 107-112.
  • Nichterlein K and Horn R(2005). Haploids in the Improvements of Linaceae and Asteraceae In: Biotechnology in Agriculture and Forestry,Palmer CE, Keller WA, Kasha, KJ (eds.), Springer- Verlag, Berlin Heidelberg Germany,pp: 286-294.
  • Nurhidayah T, Horn R, Röcher T, Friedt W(1996). High regeneration rates in anther culture of interspecific sunflower hybrids. Plant Cell Reports 16: 167-173.
  • Ozyigit II, Gozukirmizi N, Semiz BD(2007). Genotype development callus induction and shoot regeneration in sunflower (Helianthus annuus L.). Africa Journal of Biotechnology 6(13): 1498-1502.
  • Paterson KE(1984). Shoot tip culture of Helianthus annuus L.-flowering and development of adventitious and multiple shoots. American Journal of Botany 71: 925-931.
  • Pelissier B, Bouchefra O, Pepin Rand Freyssinet G(1990). Production of isolated somatic embryos from sunflower thin cell layers. Plant Cell. Rep 9: 47-50.
  • Petcu E, Pâcureanu JM(2011). Developing drought and broomrape resistant sunflower germplasm utilizing wild Helianthus species. Helia 34( 54): 1-8.
  • Phillips GC(2004). Invited review: In vitro morphogenesis in plants- Recent advances. In vitro Cell. Dev. Biol.- Plant 40: 342-345.
  • Popielarska M and PrzywaraL(2003). In vitro culture of isolated living sunflower (Helianthus annuus L.) embryo sac. Sex Plant Reprod 16: 23-33.
  • Popielarska M(2005). In vitro pollination of isolated ovules of sunflower (Helianthus annuus L.). Acta Biologica Cracoviensia Series Botanica47(1): 85–92.
  • Power CJ(1987). Organogenesis from Helianthus annuus inbreds and hybrids from the cotyledons of zygotic embryos. Am J Bot 74: 497-503.
  • Prado E and BervilleA(1990). Induction of somatic embryo development by liquid culture in Sunflower (Helianthus annuus L). Plant Sci 67: 73-82. Pugliesi C, Ceconi F, Mandolfo A, Baroncelli S(1991). Plant regeneration and genetic variability from tissue culture of Sunflower sunflower (Helianthus annuus L.). Plant Breed 106: 114-121.
  • Pugliesi C, Megale P, Cecconi F and Baroncelli S(1993). Organogenesis and embryogenesis in Helianthus tuberosus and in the interspecific hybrid Helianthus annuus X Helianthus tuberosus. Plant Cell Tissue Org Cult 33: 187-193.
  • Punia MS and Bohorova NE(1992). Callus development and plant regeneration from different explants of six wild species of sunflower (Helianthus L.). Plant Science 87: 79-83.
  • Raghavan V.(2003). One hundred years of zygotic embryo culture investigations. In vitro Cell Dev Biol-Plant 39: 437-442.
  • Rákosy-Tican E, Aurori A, Vesa S, Kovacs K-M(2007). In vitro morphogenesis of sunflower (Helianthus annuus) hypocotyl protoplasts: The effects of protoplast density, haemoglobin and spermidine. Plant Cell Tiss Organ Cult 90: 55–62.
  • Rogers MA, Gal HL, Horner HTJr, (1974). Callus formation and differentiation in tissue cultures of normal and cytoplasmic male sterile sorghum, pepper, sunflower and tobacco. In vitro 9: 463- 467.
  • Sadhu MK(1974). Effect of different auxins on growth and differentiation in callus tissue from sunflower stem pith. Indian J Exp Biol 12: 110-111.
  • Saji KV and Sujatha M(1998). Embryogenesis and plant regeneration in anther culture of sunflower (Helianthus annuus L.). Euphytica 13(1): 1-7.
  • Santos C and Caldeira G(1998). Callus formation and plant regeneration from protoplasts of sunflower calli and Hypocotyls. Acta Societatis Botanicorum Poloniae 67(1): 31-36.
  • Sarrafi A, Roustan JP, Fallot J, Alibert G(1996a). Genetic analysis of organogenesis from the cotyledons of zygotic embryos in Sunflower sunflower (Helianthus annuus L.). Theor Appl Genet 92: 225-229.
  • Sarrafi AR, BolandiH, Serieys A and AlibertG(1996b). Analysis of cotyledon culture to measure genetic variability for organogenesis parameter in sunflower (Helianthus annuus L.) Plant Sci 121: 213-219.
  • Sauca F, Lazar DA(2011). Scientific results regarding the gene(s) introgression of drought-resistance to Helianthus annuus species using embryo rescue. Romanian Biotechnological Letters 16(1): 3-8.
  • Seiler GJ(2012). Utilization of wild Helianthus species in sunflower breeding. In: Sunflower Genetics and Breeding International Monogram, ŠkorićD(ed.), Serbian Academy of Sciences and Arts, Novi Sad, Serbia,pp: 355–413.
  • Seiler GJ and Gulya T(2004). Exploration for wild Helianthus species in North America: Challenges and opportunities in the search for global treasures. Proc. 16th International Sunflower Conference, Fargo, ND USA, 43-68.
  • Seiler GJ, Rieseberg LH(1997). Systematics, Origin, and Germplasm Resources of the Wild and Domesticated Sunflower.In:Sunflower technology and production, Schneiter AA (ed.), Agron Monogr 35. ASA, CSSA and SSSA, Madison, WI, USA,p: 21-65.
  • Seiler GJ, Qi LLand Marek LF(2017). Utilization of sunflower crop wild relatives for cultivated sunflower improvement. Crop Sci 57:1083-1101.
  • Serieys H and Christov M(2005). European Cooperative Research Network on Sunflower, FAO Working Group: “Identification, Study and Utilization in Breeding Programs of New CMS Sources”, Progress Report 1999-2004, X Consultation Meeting, Novi Sad, Serbia and Montenegro, July 17-20, p:80.
  • Škorić D(1988). Sunflower breeding. J Edible Oil Ind, 25: 1-90.
  • Škorić D(2009). Sunflower breeding for resistance to abiotic stresses. Helia 32(50): 1-16.
  • Sujatha M, Vijay S, Vasavi S, Veera Reddy P, Chander RS (2012). Agrobacterium-mediated transformation of cotyledons of mature seeds of multiple genotypes of sunflower (Helianthus annuus L.). Plant Cell Tiss Organ Cult 110: 275-287.
  • Sukno S, Ruso J, Jan CC, Melero-Vara J Mand Fernandez-Martinez JM(1999). Interspecific hybridization between sunflower and wild perennial Helianthus species via embryo rescue. Euphytica106: 69–78. Tan AS, Chao-Chien Jand Gulya TJ(1992). Inheritance of resistance to Rase 4 of sunflower downy mildew in wild sunflower accessions. Crop Sci 32(4): 949-952. Thengane SR, Joshi MS, Khaspe SS and Mascarenhas AF(1994). Anther culture in Helianthus annuus L.: Influence of genotyope and culture conditions on embryo induction and plant regeneration. Plant Cell Report 13: 222- 226. Thorpe TA(1990). The current status of plant tissue culture. In: Plant Tissue Culture: Applications and Limitations, Bhojwani SS (ed.), Volume 19. 1st Edition, Elsevier, Science, Delhi, India,pp: 470.
  • Todorova M, Ivanov P, Shindrova P, Christov Mand Ivanova I(1997). Doubled haploid production of sunflower (Helianthus annuus L.) through irradiated pollen-induced parthenogenesis. Euphytica 97: 249-254.
  • Torresán A, Kesteloot J, Castano F, Rodrígues R, ColabelliM (1996). Use of immature seed germination technique as an alternative to in vitro culture of sunflower (Helianthus annuus L.) embryos. Euphytica 91: 1-3.
  • Tosun A and Ozkal N(2000). Chemical constituents and biological activities of Helianthus species. J. Fac. Pharm Ankara 29 (2): 49-74.
  • Vassilevska-Ivanova R, Shtereva L, Kraptchev B, Karceva T (2014). Response of sunflower (Helianthus annuus L) genotypes to PEG-mediated water stress. Central European Journal of Biology 9(12): 1206-1214.
  • White PR and Braun AC(1941). Crown-gall production by bacteria-free tumor tissue. Science 94(2436): 239-241.
  • Witrzens BT, Scowcroft WR, Downes RWand LarkinPJ(1988). Tissue culture and plant regeneration from sunflower (Helianthus annuus
  • L) and interspecific hybrids (H. tuberosus X H. annuus). Plant Cell Tissue Org Cult 13: 61-76.
  • Yang HY, Zhou C, Cai D, YanH, Wu Y and Chen YM(1985). In vitro culture of unfertilized ovules in Helianthus annuus L. In: Haploids of Higher Plants In vitroHu Hand Yang HY(eds), China Academic Publishers, Beijing, Springer Verlag, Berlin, pp. 182-191.
  • Yang HY, Yan H and Zhou C(1990). In Vitro Production of Haploids in Helianthus. In:Biotechnology in Agriculture and Forestry 10. Legumes and Oilseed Crops I. BajajYPS(ed), Springer-Verlag Berlin Heidelberg, pp 472-484.
  • Zhong D, MichauX-Ferriere N and Coumans M(1995). Assay for doubled haploid sunflower (Helianthus annuus) plant production by androgenesis: Fact or artifact. Part1. In vitro anther culture. Plant Cell Tissue Org Cult 41:91-100.