Optimization of the conditions for production of synthetic seeds by encapsulation of axillary buds derived from minituber sprouts in potato (Solanum tuberosum)

[EN] Synthetic seed technology is a convenient alternative to conventional multiplication in potato. In this work, we studied and optimized the process of alginate encapsulation of axillary buds derived from potato minituber sprouts (PMSs). We assayed different concentrations of sodium alginate, CaCl2 and matrix culture media, different explant sizes, different concentrations of 24-epibrassinolide (EBr) applied at different stages of the encapsulation process, different planting substrates and different cold storage periods in order to determine the best conditions for encapsulation in cvs. Sante and Agria. The conditions that resulted in the greatest regrowth rates and speed in both cultivars involved the choice of 2 3 mmlong buds, a 2 days pre-culture of buds in culture medium supplemented with 10-6 M EBr, encapsulation in 3 % sodium alginate with 1 % CaCl2 and full-strength MS culture medium, regrowth in solid MS culture medium and then transfer to coco peat (coir fiber pith) for conversion into plantlets. We also found that buds encapsulated under these conditions maintained the initial viability rates for up to 120 days in Sante and 90 days in Agria , although regrowth speed decreased after 60 days in both cultivars. We demonstrate the possibility of producing synthetic seeds efficiently using axillary buds derived from PMSs.Ghambarali, S.; Abdollahi, MR.; Zolnorian, H.; Moosavi, SS.; Seguí-Simarro, JM. (2016). Optimization of the conditions for production of synthetic seeds by encapsulation of axillary buds derived from minituber sprouts in potato (Solanum tuberosum). Plant Cell Tissue and Organ Culture (PCTOC). 126(3):449-458. doi:10.1007/s11240-016-1013-6S4494581263Adriani M, Piccioni E, Standardi A (2000) Effect of different treatments on the conversion of ‘Hayward’ kiwifruit synthetic seeds to whole plants following encapsulation of in vitro-derived buds. N Z J Crop Hortic Sci 28:59–67Ahmad N, Anis M (2010) Direct plant regeneration from encapsulated nodal segments of Vitex negundo. Biol Plant 54:748–752Azpeitia A, Chan JL, Saenz L, Oropeza C (2003) Effect of 22(S),23(S)-homobrassinolide on somatic embryogenesis in plumule explants of Cocos nucifera (L.) cultured in vitro. J Hortic Sci Biotech 78:591–596Brosa C (1999) Biological effects of brassinosteroids. Crit Rev Biochem Mol Biol 34:339–358Bustam S, Sinniah UR, Kadir MA, Zaman FQ, Subramaniam S (2012) Selection of optimal stage for protocorm-like bodies and production of artificial seeds for direct regeneration on different media and short term storage of Dendrobium Shavin White. Plant Growth Regul 69:215–224Corral-Martínez P, Seguí-Simarro JM (2014) Refining the method for eggplant microspore culture: effect of abscisic acid, epibrassinolide, polyethylene glycol, naphthaleneacetic acid, 6-benzylaminopurine and arabinogalactan proteins. Euphytica 195:369–382Divi UK, Rahman T, Krishna P (2010) Brassinosteroid-mediated stress tolerance in Arabidopsis shows interactions with abscisic acid, ethylene and salicylic acid pathways. BMC Plant Biol 10:151Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11:1–42Estrada R, Tovar P, Dodds JH (1986) Induction of in vitro tubers in a broad range of potato genotypes. Plant Cell Tissue Organ Cult 7:3–10Faisal M, Anis M (2007) Regeneration of plants from alginate-encapsulated shoots of Tylophora indica (Burm. f.) Merrill, an endangered medicinal plant. J Hortic Sci Biotechnol 82:351–354Fujioka S, Yokota T (2003) Biosynthesis and metabolism of brassinosteroids. Ann Rev Plant Biol 54:137–164Gantait S, Kundu S, Ali N, Sahu NC (2015) Synthetic seed production of medicinal plants: a review on influence of explants, encapsulation agent and matrix. Acta Physiol Plant 37:1–12Grove MD, Spencer GF, Rohwedder WK, Mandava N, Worley JF, Warthen JD, Steffens GL, Flippenanderson JL, Cook JC (1979) Brassinolide, a plant growth-promoting steroid isolated from Brassica napus pollen. Nature 281:216–217Hung CD, Trueman SJ (2012a) Alginate encapsulation of shoot tips and nodal segments for short-term storage and distribution of the eucalypt Corymbia torelliana × C. citriodora. Acta Physiol Plant 34:117–128Hung CD, Trueman SJ (2012b) Preservation of encapsulated shoot tips and nodes of the tropical hardwoods Corymbia torelliana × C. citriodora and Khaya senegalensis. Plant Cell, Tissue Organ Cult 109:341–352Jones ED (1988) A current assessment of in vitro culture and other rapid multiplication methods in North America and Europe. Am Potato J 65:209–220Kagale S, Divi UK, Krochko JE, Keller WA, Krishna P (2007) Brassinosteroid confers tolerance in Arabidopsis thaliana and Brassica napus to a range of abiotic stresses. Planta 225:353–364Kitto SL, Janick J (1982) Polyox as an artificial seed coat for asexual embryos. HortScience 17:488Larkin PJ, Davies PA, Tanner GJ (1988) Nurse culture of low numbers of Medicago and Nicotiana protoplasts using calcium alginate beads. Plant Sci 58:203–210Leclerc Y, Donnelly DJ, Coleman WK, King RR (1995) Microtuber dormancy in three potato cultivars. Am Potato J 72:215–223Machii H, Yamanouchi H (1993) Growth of mulberry synthetic seeds on vermiculite, sand and soil media. J Seric Sci Jpn 62:85–87Maguire JD (1962) Speed of germination—aid in selection ane evaluation for seedling emergence and vigor. Crop Sci 2:176–177Micheli M, Pellegrino S, Piccioni E, Standardi A (2002) Effects of double encapsulation and coating on synthetic seed conversion in M.26 apple rootstock. J Microencapsul 19:347–356Murashige T (1977) Plant cell and organ cultures as horticultural practices. Acta Hortic 78:17–30Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479Naik PS, Karihaloo JL (2007) Micropropagation for production of quality potato seed in Asia-pacific. Asia-Pacific Consortium on Agricultural Biotechnology, New DelhiNaik PS, Sarkar D (1997) Influence of light-induced greening on storage of potato microtubers. Biol Plant 39:31–34Nakashita H, Yasuda M, Nitta T, Asami T, Fujioka S, Arai Y, Sekimata K, Takatsuto S, Yamaguchi I, Yoshida S (2003) Brassinosteroid functions in a broad range of disease resistance in tobacco and rice. Plant J 33:887–898Nassar KAM, Kubow S, Donnelly JD (2015) Somatic Embryogenesis for potato (Solanum tuberosum L.) improvement. In: Li X-Q, Donnelly JD, Jensen GT (eds) Somatic genome manipulation: advances, methods, and applications. Springer, New York, pp 169–197Nuñez M, Siqueira WJ, Hernandez M, Zullo MAT, Robaina C, Coll F (2004) Effect of spirostane analogues of brassinosteroids on callus formation and plant regeneration in lettuce (Lactuca sativa). Plant Cell Tissue Organ Cult 78:97–99Nyende AB, Schittenhelm S, Mix-Wagner G, Greef J-M (2003) Production, storability, and regeneration of shoot tips of potato (Solanum tuberosum L.) encapsulated in calcium alginate hollow beads. In Vitro Cell Dev Biol Plant 39:540–544Oh MH, Clouse SD (1998) Brassinolide affects the rate of cell division in isolated leaf protoplasts of Petunia hybrida. Plant Cell Rep 17:921–924Parveen S, Shahzad A (2014) Encapsulation of nodal segments of Cassia angustifolia Vahl. for short-term storage and germplasm exchange. Acta Physiol Plant 36(3):635–640Pullman GS, Zhang Y, Phan BH (2003) Brassinolide improves embryogenic tissue initiation in conifers and rice. Plant Cell Rep 22:96–104Rai MK, Asthana P, Singh SK, Jaiswal VS, Jaiswal U (2009) The encapsulation technology in fruit plants—a review. Biotechnol Adv 27:671–679Redenbaugh K, Paasch BD, Nichol JW, Kossler ME, Viss PR, Walker KA (1986) Somatic seeds: encapsulation of asexual plant embryos. Nat Biotechnol 4:797–801Rihan HZ, Al-Issawi M, Burchett S, Fuller MP (2011) Encapsulation of cauliflower (Brassica oleracea var botrytis) microshoots as artificial seeds and their conversion and growth in commercial substrates. Plant Cell Tissue Organ Cult 107:243–250Saha S, Sengupta C, Ghosh P (2015) Encapsulation, short-term storage, conservation and molecular analysis to assess genetic stability in alginate-encapsulated microshoots of Ocimum kilimandscharicum Guerke. Plant Cell Tissue Organ Cult 120:519–530Sarkar D, Naik PS (1997) Nutrient-encapsulation of potato nodal segments for germplasm exchange and distribution. Biol Plant 40:285–290Sarkar D, Naik PS (1998) Synseeds in potato: an investigation using nutrient-encapsulated in vitro nodal segments. Sci Hortic 73:179–184Sasaki H (2002) Brassinolide promotes adventitious shoot regeneration from cauliflower hypocotyl segments. Plant Cell Tissue Organ Cult 71:111–116Schafer-Menuhr A, Mix-Wagner G, Vorlop K (2003) Regeneration of plants from cell suspension cultures and encapsulated cell suspension cultures of Solanum tuberosum L. cv. Clarissa. Landbauforsch Völkenrode 53:53–59Sharma S, Shahzad A (2012) Encapsulation technology for short-term storage and conservation of a woody climber, Decalepis hamiltonii Wight and Arn. Plant Cell Tissue Organ Cult 111:191–198Sharma SK, Bryan GJ, Winfield MO, Millam S (2007) Stability of potato (Solanum tuberosum L.) plants regenerated via somatic embryos, axillary bud proliferated shoots, microtubers and true potato seeds: a comparative phenotypic, cytogenetic and molecular assessment. 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Production of doubled haploid plants from anther cultures of borage (Borago officinalis L.) by the application of chemical and physical stress

[EN] Anther culture can be used as a powerful tool to produce doubled haploid (DH) lines in medicinal plants, thus accelerating breeding programs. In the particular case of borage (Borago officinalis L.), a method to produce DH plants has not been yet published. In this work we evaluated the effect of different culture media and of different chemical (colchicine and n-butanol) and physical stresses (centrifugation and electroporation) on androgenesis induction and plant regeneration in borage anther cultures. We found that the highest response can be obtained with culture medium containing B5 salts and NLN vitamins, the addition of 200 mg/l colchicine during 4 days, a pretreatment of anthers with 0.2% n-butanol for 5 hours, or the application to anthers of single physical stresses (either centrifugation at 300 g or a 100 v electrical shock, but not combined). This is the first report on the production of DH plants in borage. Together, the results presented hereby can be used as a basic framework for large-scale generation of DH plants in this important medicinal species.Hoveida, Z.; Abdollahi, MR.; Mirzaie-Asl, A.; Moosavi, SS.; Seguí-Simarro, JM. (2017). Production of doubled haploid plants from anther cultures of borage (Borago officinalis L.) by the application of chemical and physical stress. Plant Cell Tissue and Organ Culture (PCTOC). 130(2):369-378. doi:10.1007/s11240-017-1233-4S3693781302Alemanno L, Guiderdoni E (1994) Increased doubled haploid plant regeneration from rice (Oryza sativa L.) anthers cultured on colchicine-supplemented media. Plant Cell Rep 13:432–436Bayliss K, Wroth J, Cowling W (2004) Pro-embryos of Lupinus spp. produced from isolated microspore culture. Crop Pasture Sci 55:589–593Castillo AM, Cistué L, Vallés MP, Soriano M (2009) Chromosome doubling in monocots. In: Touraev A, Forster BP, Jain SM (eds) Advances in haploid production in higher plants. Springer, Dordrecht, pp 329–338Castillo AM, Nielsen NH, Jensen A, Vallés MP (2014) Effects of n-butanol on barley microspore embryogenesis. Plant Cell Tissue Organ Cult 117:411–418Chardoli Eshaghi Z, Abdollahi MR, Moosavi SS, Deljou A, Seguí-Simarro JM (2015) Induction of androgenesis and production of haploid embryos in anther cultures of borage (Borago officinalis L.). Plant Cell Tissue Organ Cult 122(2):1–9Chu CC (1978) The N6 medium and its applications to anther culture of cereal crops. In: Proceedings of symposium on plant tissue culture, 25–30 May 1978. Science Press, Beijing, pp 45–50Das UN (2006) Tumoricidal and anti-angiogenic actions of gamma-linolenic acid and its derivatives. Curr Pharm Biotechnol 7:457–466De Lisi A, Montesano V, Negro D, Sarli G, Blanco E, Sonnante G, Laghetti G (2014) Genetic diversity in Borago officinalis germplasm as revealed by seed oils and AFLP polymorphism. Genet Resour Crop Evol 61:853–859Delaitre C, Ochatt S, Deleury E (2001) Electroporation modulates the embryogénie responses of asparagus (Asparagus officinalis L.) microspores. Protoplasma 216:39–46Fábián A, Földesiné Füredi PK, Ambrus H, Jäger K, Szabó L, Barnabás B (2015) Effect of n-butanol and cold pretreatment on the cytoskeleton and the ultrastructure of maize microspores when cultured in vitro. Plant Cell Tissue Organ Cult 123:257–271Ferrie AMR (2007) Doubled haploid production in nutraceutical species: a review. Euphytica 158:347–357Ferrie AMR (2009) Current status of doubled haploids in medicinal plants. Advances in haploid production in higher plants. Springer, Dordrecht, pp 209–217Ferrie AMR (2013) Advances in microspore culture technology: a biotechnological tool for the improvement of medicinal plants. In: Chandra S, Lata H, Varma A (eds) Biotechnology for medicinal plants: micropropagation and improvement. Springer, Berlin, pp 191–206Ferrie AMR, Bethune TD, Mykytyshyn M (2011) Microspore embryogenesis in Apiaceae. Plant Cell Tissue Organ Cult 104:399–406Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158Germanà MA (2011) Anther culture for haploid and doubled haploid production. Plant Cell Tissue Organ Cult 104:283–300Grewal RK, Lulsdorf M, Croser J, Ochatt S, Vandenberg A, Warkentin TD (2009) Doubled-haploid production in chickpea (Cicer arietinum L.): role of stress treatments. Plant Cell Rep 28:1289–1299Kapoor R, Huang YS (2006) Gamma linolenic acid: an antiinflammatory omega-6 fatty acid. Curr Pharm Biotechnol 7:531–534Keller WA, Armstrong KC (1977) Embryogenesis and plant regeneration in Brassica napus anther cultures. Can J Bot 55:1383–1388Lichter R (1982) Induction of haploid plants from isolated pollen of Brassica napus. Z Pflanzenphysiol 105: 427–434Lulsdorf MM, Croser JS, Ochatt S (2011) Androgenesis and doubled-haploid production in food legumes. In Pratap A, Kumar J (eds) Biology and breeding of food legumes. CABI Publishers, Wallingford, pp 159–177Montaner C, Floris E, Alvarez JM (2000) Is self-compatibility the main breeding system in borage (Borago officinalis L.)? Theor Appl Genet 101:185–189Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479Nitsch JP, Nitsch C (1969) Haploid plants from pollen grains. Science 163:85–87Obert B, Barnabás B (2004) Colchicine induced embryogenesis in maize. Plant Cell Tissue Organ Cult 77:283–285Ochatt S (2013) Plant cell electrophysiology: applications in growth enhancement, somatic hybridisation and gene transfer. Biotechnol Adv 31:1237–1246Ochatt S, Pech C, Grewal R, Conreux C, Lulsdorf M, Jacas L (2009) Abiotic stress enhances androgenesis from isolated microspores of some legume species (Fabaceae). J Plant Physiol 166:1314–1328Pathirana R, Frew T, Hedderley D, Timmerman-Vaughan G, Morgan E (2011) Haploid and doubled haploid plants from developing male and female gametes of Gentiana triflora. Plant Cell Rep 30:1055–1065Phillips GC, Collins GB (1979) In vitro tissue culture of selected legumes and plant regeneration from callus cultures of red clover. Crop Sci 19:59–64Ribalta FM, Croser JS, Ochatt SJ (2012) Flow cytometry enables identification of sporophytic eliciting stress treatments in gametic cells. J Plant Physiol 169:104–110Sales E, Montaner C, Muniozguren JM, Carravedo M, Alvarez JM (2008) Genetic diversity in a collection of borage (Borago officinalis) germplasm. Botany 86:603–609Seguí-Simarro JM (2010) Androgenesis revisited. Bot Rev 76:377–404Seguí-Simarro JM, Nuez F (2007) Embryogenesis induction, callogenesis, and plant regeneration by in vitro culture of tomato isolated microspores and whole anthers. J Exp Bot 58:1119–1132Seguí-Simarro JM, Nuez F (2008) Pathways to doubled haploidy: chromosome doubling during androgenesis. Cytogenet Genome Res 120:358–369Shariatpanahi ME, Bal U, Heberle-Bors E, Touraev A (2006) Stresses applied for the re-programming of plant microspores towards in vitro embryogenesis. Physiol Plant 127:519–534Soriano M, Cistué L, Vallés MP, Castillo AM (2007) Effects of colchicine on anther and microspore culture of bread wheat (Triticum aestivum L.). Plant Cell Tissue Organ Cult 91:225–234Soriano M, Cistue L, Castillo AM (2008) Enhanced induction of microspore embryogenesis after n-butanol treatment in wheat (Triticum aestivum L.) anther culture. Plant Cell Rep 27:805–811Szakács É, Barnabás B (1995) The effect of colchicine treatment on microspore division and microspore-derived embryo differentiation in wheat (Triticum aestivum L.) anther culture. Euphytica 83:209–213Tanaka M (1973) The effect of centrifugal treatment on the emergence of plantlet from cultured anther of tobacco. Japan J Breed 23: 171–174Touraev A, Ilham A, Vicente O, Heberle-Bors E (1996) Stress-induced microspore embryogenesis in tobacco: an optimized system for molecular studies. Plant Cell Rep 15:561–565Uno Y, Koda-Katayama H, Kobayashi H (2016) Application of anther culture for efficient haploid production in the genus Saintpaulia. Plant Cell Tissue Organ Cult 125:241–248Zhou WJ, Tang GX, Hagberg P (2002) Efficient production of doubled haploid plants by immediate colchicine treatment of isolated microspores in winter Brassica napus. Plant Growth Regul 37:185–19

Induction of androgenesis and production of haploid embryos in anther cultures of borage (Borago officinalis L.)

[EN] Borage (Borago officinalis L.) is an important medicinal plant with different culinary, pharmaceutical and industrial properties. Unfortunately, there are no published reports on the establishment of protocols to produce DHs in this species up to now. In this work, we show for the first time the induction of borage microspores to become embryogenic calli, from which haploid embryos are produced. In addition, we evaluated the effect of using different flower bud sizes, carbon sources, concentrations of 2,4-D and BAP, cold (4 A degrees C) pretreatments and heat shock treatments. Production of total calli, embryogenic calli and callus-derived embryos was differently affected by the different parameters studied. Our results showed that the use of 5-7 mm-long flower buds, a cold (4 A degrees C) pretreatment during 4 days, a 32 A degrees C heat shock for 3 days, and the addition of 3 % maltose and 2 mgl(-1) 2,4-D and 1 mgl(-1) BAP to the culture medium, was beneficial for embryo production. Overall, this work demonstrates that DH technology is possible in borage, and opens the door for future improvements needed to finally obtain borage DH plants.Eshaghi, ZC.; Abdollahi, MR.; Moosavi, SS.; Deljou, A.; Seguí-Simarro, JM. (2015). Induction of androgenesis and production of haploid embryos in anther cultures of borage (Borago officinalis L.). Plant Cell, Tissue and Organ Culture. 122:321-329. doi:10.1007/s11240-015-0768-5S321329122Abdollahi MR, Moieni A, Javaran MJ (2004) Interactive effects of shock and culture density on embryo induction in isolated microspore culture of Brassica napus L. cv. Global Iranian J Biotech 2:97–100Bohanec B, Neskovic M, Vujicic R (1993) Anther culture and androgenetic plant regeneration in buckwheat (Fagopyrum esculentum Moench). Plant Cell Tissue Organ Cult 35:259–266Calleberg E, Johansson L (1996) Effect of gelling agents on anther cultures. In: Jain SM, Sopory SK, Veilleux RE (eds) In vitro haploid production in higher plants, vol 23. Springer, Netherlands, pp 189–203Custers JBM, Cordewener JHG, Nöllen Y, Dons JJ, van Lookeren-Campagne MM (1994) Temperature controls both gametophytic and sporophytic development in microspore cultures of Brassica napus. Plant Cell Rep 13:267–271Ferrie AMR (2013) Advances in microspore culture technology: a biotechnological tool for the improvement of medicinal plants. In: Chandra S et al (eds) Biotechnology for medicinal plants. Springer, Berlin, pp 191–206Ferrie AMR, Caswell KL (2011) Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production. Plant Cell Tissue Organ Cult 104:301–309Ferrie AMR, Bethune T, Mykytyshyn M (2011) Microspore embryogenesis in the Apiaceae. Plant Cell Tissue Organ Cult 104:399–406Forster BP, Heberle-Bors E, Kasha KJ, Touraev A (2007) The resurgence of haploids in higher plants. Trends Plant Sci 12:368–375Gamborg OL, Miller RA, Ojiwa K (1968) Nutrient requirements of suspension culture of soybean root callus. Exp Cell Res 50:151–158Guil-Guerrero JL, García-Maroto F, Vilches-Ferrón MA, López-Alonso D (2003) Gamma-linolenic acid from fourteen Boraginaceae species. Ind Crop Prod 18:85–89Horrobin DF (1983) The regulation of prostaglandin biosynthesis by the manipulation of essential fatty acid metabolism. Rev Pure Appl Pharmacol Sci 4:339–383Irikova T, Grozeva S, Rodeva V (2011) Anther culture in pepper (Capsicum annuum L.) in vitro. Acta Physiol Plant 33:1559–1570Lauxen MS, Kaltchuk-Santos E, Hu CY, Callegari-Jacques SM, Bodanese-Zanettini MH (2003) Association between floral bud size and developmental stage in soybean microspores. Braz Arch Biol Technol 46:515–520Leach CR, Mayo O, Bürger R (1990) Quantitatively determined self-incompatibility. Outcrossing in Borago officinalis. Theoret Appl Genetics 79:427–430Lichter R (1982) Induction of haploid plants from isolated pollen of Brassica napus. Z Pflanzenphysiol 105:427–434Maluszynski M, Kasha KJ, Szarejko I (2003) Published doubled haploid protocols in plant species. In: Maluszynski M, Kasha KJ, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants. A manual. Kluwer, Dordrecht, pp 309–335Maraschin SF, de Priester W, Spaink HP, Wang M (2005) Androgenic switch: an example of plant embryogenesis from the male gametophyte perspective. J Exp Bot 56:1711–1726McDonald BE, Fitzpatrick K (1998) Designer Vegetable Oils. In: Mazza G (ed) Functional foods, biochemical and processing aspects. Technomic Publ Co. Inc, Lancaster, pp 265–291Ozkum D, Tipirdamaz R (2002) The effects of cold treatment and charcoal on the in vitro androgenesis of pepper (Capsicum annuum L.). Turk J Bot 26:131–139Parra-Vega V, González-García B, Seguí-Simarro JM (2013a) Morphological markers to correlate bud and anther development with microsporogenesis and microgametogenesis in pepper (Capsicum annuum L.). Acta Physiol Plant 35:627–633Parra-Vega V, Renau-Morata B, Sifres A, Seguí-Simarro JM (2013b) Stress treatments and in vitro culture conditions influence microspore embryogenesis and growth of callus from anther walls of sweet pepper (Capsicum annuum L.). Plant Cell Tissue Organ Cult 112:353–360Raquin C (1983) Utilization of different sugars as carbon sources for in vitro cultures of Petuina. Z Pflanzenphysol 111:453–457Salas P, Rivas-Sendra A, Prohens J, Seguí-Simarro JM (2012) Influence of the stage for anther excision and heterostyly in embryogenesis induction from eggplant anther cultures. Euphytica 184:235–250Seguí-Simarro JM (2010) Androgenesis revisited. Bot Rev 76:377–404Seguí-Simarro JM, Nuez F (2006) Androgenesis induction from tomato anther cultures: callus characterization. Acta Hort 725:855–861Seguí-Simarro JM, Corral-Martínez P, Parra-Vega V, González-García B (2011) Androgenesis in recalcitrant solanaceous crops. Plant Cell Rep 30:765–778Shariatpanahi ME, Bal U, Heberle-Bors E, Touraev A (2006) Stresses applied for the reprogramming of plant microspores towards in vitro embryogenesis. Physiol Plant 127:519–534Simon JE, Chadwick AF, Craker LE (1984) Herbs: an indexed bibliography. 1971–1980. The scientific literature on selected herbs, and aromatic and medicinal plants of the temperate zone. Archon Books, Hamden, CTSkrzypek E, Czyczyło-Mysza I, Marcińska I, Wędzony M (2008) Prospects of androgenetic induction in Lupinus spp. Plant Cell Tissue Organ Cult 94(2):131–137Snape JW (1989) Doubled haploid breeding: theoretical basis and practical applications. In: Mujeeb-Kazi A, Sitch LA (eds) Review of advances in plant biotechnology, 1985–1988: 2nd international symposium genetic manipulation in crops. Mexico and Manila, CIMMYT and IRRI, pp 19–30Tipirdamaz R, Ellialtioğlu Ş (1998) The effects of cold treatments and activated charcoal on ABA contents of anthers and in vitro androgenesis in eggplant (Solanum melongena L.). In: Tsekos I, Moustakas M (eds) Progress in botanical research, Proceedings of the 1st Balkan botanical congress. Kluwer Academic Publishers, The NetherlandsVagera J, Havranek P (1985) In vitro induction of androgenesis in Capsicum annuum L. and its genetic aspests. Biol Plant 27(1):10–21Zur I, Dubas E, Golemiec E, Szechynska-Hebda M, Golebiowska G, Wedzony M (2009) Stress-related variation in antioxidative enzymes activity and cell metabolism efficiency associated with embryogenesis induction in isolated microspore culture of triticale (×Triticosecale Wittm.). Plant Cell Rep 28:1279–128

Chloroplastic acyl carrier protein synthase I and chloroplastic 20 kDa chaperonin proteins are involved in wheat (Triticum aestivum) in response to moisture stress

In this study, two bread wheat (Triticum aestivum L.) cultivars, Pishgam (drought-tolerant) and Shahryar (drought-sensitive), were grown in the greenhouse under control and moisture stress conditions. Based on phenological and morpho-physiological results, Pishgam was confirmed as a moisture stress tolerant cultivar. In the fallowing, at the start of heading time, its treated and untreated flag leaves were sampled for two-dimensional electrophoresis (2-DE) based on proteomics approach. Among approximately 263 protein spots appearing in two-dimensional gels, 23 and 10 protein spots were up- and down-regulated, respectively. Among these differentially expressed proteins, 11 proteins with more differences were identified by MALDI TOF/TOF MS which allocated to six functional protein groups involved in photosynthesis or respiration, carbohydrate metabolism, energy metabolism, chaperon, lipid metabolism and unknown function. We report this for the first time that chloroplastic acyl carrier protein synthase I and chloroplastic 20 kDa chaperonin proteins were significantly changed in wheat in response to moisture stress