MORPHO-HISTOLOGICAL ASPECTS OF ADVENTITIOUS SHOOT FORMATION WITHOUT PLANT GROWTH REGULATORS IN SEED EXPLANTS OF Capsicum annuum L., AND IMPACT OF PRECULTURE ON REGENERATION
Andrzej Gatz
University of Technology and Life Sciences in BydgoszczUniversity of Technology and Life Sciences in BydgoszczAbstract
The physiological state of plant material is the crucial endogenous factor at the explant choice for plant regeneration. The phases of germination characterised by various, following each other biochemical and developmental processes can affect the organogenesis capability. This research examined the morphological and anatomical events during the early stages of organogenesis and plant regeneration in explants derived from seeds of Capsicum annuum L., cv. Bryza preincubated under high humidity conditions from 0 to 6 days and next cultured on MS medium without PGRs. The early stage of de novo shoot formation reminded leaf differentiation in planta. First the leaves began to differentiate as spherical and tongue shaped structures from epidermis and subjacent layers of the explants about the 7th day of culture. In some cases nearly at the base of previously formed leaf and even on its petiole one or two leaves as well as shoot apex in their axils
were induced thereby forming young shoot which underwent elongation and whole plant regeneration after 2 subculturing. More advanced developmentally structures of adventitious shoot were obtained while prolonging preculture duration. This was the favourable effect on the shoot differentiation, their elongation and plant regeneration as seed submitted preculture for 3, 4, 5 days however, seeds not treated with preculture revealed the best response as regard to shoot primordium formation at the earliest stage.
Keywords:
pepper, shoot morphogenesis, germination, histogenesis, leaf formation in vitro, plant regeneration without PGRs, shoot apical meristemReferences
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Blasiak J., Kuang A., Farhangi C.S., Musgrave M.E., 2006. Roles of intra-fruit oxygen and carbon dioxide in controlling pepper (Capsicum annuum L.) seed development and storage reserve deposition. J. Amer. Soc. Hort. Sci 131, 164–173.
Bowes B.G., 1976. Polar regeneration in excised roots of Taraxacum officinale weber: a light and electron microscopic study. Ann. Bot. 40, 423–432.
Darlington G.D., La Cour L.F., 1960. The handling of chromosomes. London. Unwin Ltd. 163.
De Klerk G.J., Arnholdt-Schmitt B., Lieberei R., Neumann K.H., 1997. Regeneration of roots, shoots and embryos: physiological, biochemical and molecular aspects. Biol. Plant. 39, 53–66.
Delis M., Garbaczewska G., Niemirowicz-Szczytt K., 2005. Differentiation of adventitious buds from Capsicum annuum L. hypocotyls after co-culture with Agrobacterium tumefaciens. Acta Biol. Cracov. Ser. Bot. 47, 193–198.
Diaz I., Moreno R., Power J.B., 1988. Plant regeneration from protoplasts of Capsicum annuum. Plant Cell Rep. 7, 210–212.
Duclercq J., Sangwan-Norrel B., Catterou M., Sangwan R.S, 2011. De novo shoot organogenesis: from art to science. Trends Plant Sci. 16, 597–606.
Dun E.A., Brewer P.B., Beveridge C.A., 2009. Strigolactones: discovery of the elusive shoot branching hormone. Trends Plant Sci. 14, 364–372.
Ebida A.I.A., Hu C.Y., 1993. In vitro morphogenetic responses and plant regeneration from pepper (Capsicum annuum L. cv. Early California Wonder) seedling explants. Plant Cell Rep. 13, 107–110.
Elhiti M., Stasolla C., 2012. In vitro shoot organogenesis and hormone response are affected by the altered levels of Brassica napus meristem genes. Plant Sci. 190, 40–51.
Ezura H., Nishimiya S., Kasumi M., 1993. Efficient regeneration of plants independent of exogenous growth regulators in bell pepper (Capsicum annuum L.). Plant Cell. Rep. 12, 676–680.
Fari M., Czako M., 1981. Relationship between position and morphogenetic response of pepper hypocotyl explant cultured in vitro. Sci. Hortic. Amsterdam 15, 207–213
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Gatz A., Rogozińska J., 1994. In vitro organogenetic potential of cotyledon and leaf explants of Capsicum annuum L. cv. Bryza. Acta Soc. Bot. Pol. 63, 255–258.
Gatz A., 2002. Histological changes during the adventitious shoot formation in seedling explants of pepper (Capsicum annuum L.) cultured in vitro. Acta Soc. Bot. Pol. 71, 269–274.
George L., Narayanaswamy S., 1973. Haploid Capsicum through experimental androgenesis. Protoplasma 78, 467–470.
Gunay A.L., Rao P.S., 1978. In vitro plant regeneration from hypocotyl and cotyledon explants of red pepper (Capsicum). Plant Sci. Lett. 11, 365–372.
Harini I., Lakshmi Sita G., 1993. Direct somatic embryogenesis and plant regeneration from immature embryos of chilli (Capsicum annuum L.). Plant Sci. 89, 107–112.
Harper L., Freeling M., 1996. Studies on early leaf development. Curr. Opin. Plant Biot. 7, 139–144.
Imaichi R., Hiyama Y., Kato M., 2005. Leaf development in the absence of a shoot apical meristem in Zeylanidium subulatum (Podostemacea). Ann. Bot. 96, 51–58.
Keller T., Abbott J., Moritz T., Doerner P., 2006. Arabidopsis REGULATOR OF AXILLARY MERISTEMS1 controls a leaf axil stem cell niche and modulates vegetative development. Plant Cell. 18, 598–611.
Khan H., Siddique I., Anis M., 2006. Thidiazuron induced somatic embryogenesis and plant regeneration in Capsicum annuum. Biol. Plant 50, 789–792.
Kothari S.L., Joshi A., Kachhwaha S., Ochoa-Alejo N., 2010. Chili peppers. A review on tissue culture and transgenesis. Biotech. Adv. 28, 35–48.
Lijun O., Zhuqing Z., Xiongze D., Xuexiao Z., 2012. Seed micromorphology and germination characteristics of wild and cultivated pepper strains. Plant Syst. Evol. 298, 1211–1216.
Ljung K., Ostin A., Lioussanne L., Sandberg G., 2001. Developmental regulation of indole-3-acetic acid turnover in Scot pine seedlings. Plant Physiol. 125, 464–475.
Mac Connell J.R., Barton M.K., 1995. Effect of mutations in the PINHEAD gene of Arabidopsis on the formation of shoot apical meristems. Dev. Genet. 16, 358–366.
Mezghani N., Jemmali A., Elloumi N., Gargouri-Bouzid R., Kintzios S., 2007. Morphohistological study on shoot bud regeneration in cotyledon cultures of pepper (Capsicum annuum). Biologia 62, 704–710.
Murashige T., Skoog F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiol. 15, 473–497.
Ochoa-Alejo N., Ireta-Moreno L., 1990. Cultivar differences in shoot-forming capacity of hypocotyls tissues of chili pepper (Capsicum annuum) cultured in vitro. Sci. Hortic. 42, 261–269.
Nonogaki H., Bassel G.W., Bewley J.D., 2010. Germination – still a mystery. Plant Sci., 179, 574–581.
Phillips G.C., Hubstenberger J.F., 1985. Organogenesis in pepper tissue cultures. Plant Cell. Tiss. Org. Cult. 4, 261–269.
Pierik R.L.M., 1987. In vitro culture of higher plants. Martinus Nijhoff Publishers Dordrecht. Ramirez-Malagon R., Ochoa-Alejo N., 1996. An improved and reliable chili pepper (Capsicum annuum L.) plant regeneration method. Plant Cell. Rep. 16, 226–231.
Ruyter-Spira C., Al-Babili S., Van der Krol S., Bouwmeester H., 2013. The biology of strigolactones. Trends Plant Sci. 18, 72–83.
Sattler R., Maier U., 1977. Development of epiphyllous appendages of Begonia hispida var. cucullifera – implications for comparative morphology. Can. J. Bot. 55, 411–425.
Saxena P.K., Gill R., Rashid A., Maheshwari S.C., 1981. Isolation and culture of protoplasts of Capsicum annuum L. and their regeneration into plants flowering in vitro. Protoplasma 108, 357–360.
Scanlon J.M., 2000. Developmental complexities of simple leaves. Curr. Opin. Plant Biol. 3, 31–36.
Selker J.M.L., Lyndon R.F., 1996. Leaf initiation and de novo pattern formation in the absence of an apical meristem and pre-existing patterned leaves in watercress (Nasturtium officinale) axillary explants. Can. J. Bot.74, 625–641.
Segui-Simarro J.M., Corral-Martinez P., Parra-Vega V., Gonzalez-Garcia B., 2011. Androgenesis in recalcitrant solanaceous crops. Plant Cell. Rep. 30, 765–778.
Sripichitt P., Nawata E., Shigenaga S., 1988. The effect of exposure dose and dose rate of gamma radiation on in vitro shoot forming capacity of cotyledon explants in red pepper (Capsicum annuum L. cv. Yatsufusa). Jap. J. Breed. 38, 27–34.
Su Y.H., Liu Y.B., Zhang X.S., 2011. Auxin-cytokinin interaction regulates meristem development. Mol. Plant 4(4), 616–625.
Szasz A., Nervo G., Fari M., 1995. Screening for in vitro shoot-forming capacity of seedling explants in bell pepper (Capsicum annuum L.) genotypes and efficient plant regeneration using thidiazuron. Plant Cell. Rep. 14, 666–669.
Blasiak J., Kuang A., Farhangi C.S., Musgrave M.E., 2006. Roles of intra-fruit oxygen and carbon dioxide in controlling pepper (Capsicum annuum L.) seed development and storage reserve deposition. J. Amer. Soc. Hort. Sci 131, 164–173.
Bowes B.G., 1976. Polar regeneration in excised roots of Taraxacum officinale weber: a light and electron microscopic study. Ann. Bot. 40, 423–432.
Darlington G.D., La Cour L.F., 1960. The handling of chromosomes. London. Unwin Ltd. 163.
De Klerk G.J., Arnholdt-Schmitt B., Lieberei R., Neumann K.H., 1997. Regeneration of roots, shoots and embryos: physiological, biochemical and molecular aspects. Biol. Plant. 39, 53–66.
Delis M., Garbaczewska G., Niemirowicz-Szczytt K., 2005. Differentiation of adventitious buds from Capsicum annuum L. hypocotyls after co-culture with Agrobacterium tumefaciens. Acta Biol. Cracov. Ser. Bot. 47, 193–198.
Diaz I., Moreno R., Power J.B., 1988. Plant regeneration from protoplasts of Capsicum annuum. Plant Cell Rep. 7, 210–212.
Duclercq J., Sangwan-Norrel B., Catterou M., Sangwan R.S, 2011. De novo shoot organogenesis: from art to science. Trends Plant Sci. 16, 597–606.
Dun E.A., Brewer P.B., Beveridge C.A., 2009. Strigolactones: discovery of the elusive shoot branching hormone. Trends Plant Sci. 14, 364–372.
Ebida A.I.A., Hu C.Y., 1993. In vitro morphogenetic responses and plant regeneration from pepper (Capsicum annuum L. cv. Early California Wonder) seedling explants. Plant Cell Rep. 13, 107–110.
Elhiti M., Stasolla C., 2012. In vitro shoot organogenesis and hormone response are affected by the altered levels of Brassica napus meristem genes. Plant Sci. 190, 40–51.
Ezura H., Nishimiya S., Kasumi M., 1993. Efficient regeneration of plants independent of exogenous growth regulators in bell pepper (Capsicum annuum L.). Plant Cell. Rep. 12, 676–680.
Fari M., Czako M., 1981. Relationship between position and morphogenetic response of pepper hypocotyl explant cultured in vitro. Sci. Hortic. Amsterdam 15, 207–213
Fari M., 1986. Pepper (Capsicum annuum L.). In: Biotechnology in Agriculture and Forestry, vol. 2, Crops, Bajaj Y.P.S. (ed.). Springer Verlag Berlin, Heidelberg, New York, Tokyo 345–362.
Gatz A., Rogozińska J., 1994. In vitro organogenetic potential of cotyledon and leaf explants of Capsicum annuum L. cv. Bryza. Acta Soc. Bot. Pol. 63, 255–258.
Gatz A., 2002. Histological changes during the adventitious shoot formation in seedling explants of pepper (Capsicum annuum L.) cultured in vitro. Acta Soc. Bot. Pol. 71, 269–274.
George L., Narayanaswamy S., 1973. Haploid Capsicum through experimental androgenesis. Protoplasma 78, 467–470.
Gunay A.L., Rao P.S., 1978. In vitro plant regeneration from hypocotyl and cotyledon explants of red pepper (Capsicum). Plant Sci. Lett. 11, 365–372.
Harini I., Lakshmi Sita G., 1993. Direct somatic embryogenesis and plant regeneration from immature embryos of chilli (Capsicum annuum L.). Plant Sci. 89, 107–112.
Harper L., Freeling M., 1996. Studies on early leaf development. Curr. Opin. Plant Biot. 7, 139–144.
Imaichi R., Hiyama Y., Kato M., 2005. Leaf development in the absence of a shoot apical meristem in Zeylanidium subulatum (Podostemacea). Ann. Bot. 96, 51–58.
Keller T., Abbott J., Moritz T., Doerner P., 2006. Arabidopsis REGULATOR OF AXILLARY MERISTEMS1 controls a leaf axil stem cell niche and modulates vegetative development. Plant Cell. 18, 598–611.
Khan H., Siddique I., Anis M., 2006. Thidiazuron induced somatic embryogenesis and plant regeneration in Capsicum annuum. Biol. Plant 50, 789–792.
Kothari S.L., Joshi A., Kachhwaha S., Ochoa-Alejo N., 2010. Chili peppers. A review on tissue culture and transgenesis. Biotech. Adv. 28, 35–48.
Lijun O., Zhuqing Z., Xiongze D., Xuexiao Z., 2012. Seed micromorphology and germination characteristics of wild and cultivated pepper strains. Plant Syst. Evol. 298, 1211–1216.
Ljung K., Ostin A., Lioussanne L., Sandberg G., 2001. Developmental regulation of indole-3-acetic acid turnover in Scot pine seedlings. Plant Physiol. 125, 464–475.
Mac Connell J.R., Barton M.K., 1995. Effect of mutations in the PINHEAD gene of Arabidopsis on the formation of shoot apical meristems. Dev. Genet. 16, 358–366.
Mezghani N., Jemmali A., Elloumi N., Gargouri-Bouzid R., Kintzios S., 2007. Morphohistological study on shoot bud regeneration in cotyledon cultures of pepper (Capsicum annuum). Biologia 62, 704–710.
Murashige T., Skoog F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiol. 15, 473–497.
Ochoa-Alejo N., Ireta-Moreno L., 1990. Cultivar differences in shoot-forming capacity of hypocotyls tissues of chili pepper (Capsicum annuum) cultured in vitro. Sci. Hortic. 42, 261–269.
Nonogaki H., Bassel G.W., Bewley J.D., 2010. Germination – still a mystery. Plant Sci., 179, 574–581.
Phillips G.C., Hubstenberger J.F., 1985. Organogenesis in pepper tissue cultures. Plant Cell. Tiss. Org. Cult. 4, 261–269.
Pierik R.L.M., 1987. In vitro culture of higher plants. Martinus Nijhoff Publishers Dordrecht. Ramirez-Malagon R., Ochoa-Alejo N., 1996. An improved and reliable chili pepper (Capsicum annuum L.) plant regeneration method. Plant Cell. Rep. 16, 226–231.
Ruyter-Spira C., Al-Babili S., Van der Krol S., Bouwmeester H., 2013. The biology of strigolactones. Trends Plant Sci. 18, 72–83.
Sattler R., Maier U., 1977. Development of epiphyllous appendages of Begonia hispida var. cucullifera – implications for comparative morphology. Can. J. Bot. 55, 411–425.
Saxena P.K., Gill R., Rashid A., Maheshwari S.C., 1981. Isolation and culture of protoplasts of Capsicum annuum L. and their regeneration into plants flowering in vitro. Protoplasma 108, 357–360.
Scanlon J.M., 2000. Developmental complexities of simple leaves. Curr. Opin. Plant Biol. 3, 31–36.
Selker J.M.L., Lyndon R.F., 1996. Leaf initiation and de novo pattern formation in the absence of an apical meristem and pre-existing patterned leaves in watercress (Nasturtium officinale) axillary explants. Can. J. Bot.74, 625–641.
Segui-Simarro J.M., Corral-Martinez P., Parra-Vega V., Gonzalez-Garcia B., 2011. Androgenesis in recalcitrant solanaceous crops. Plant Cell. Rep. 30, 765–778.
Sripichitt P., Nawata E., Shigenaga S., 1988. The effect of exposure dose and dose rate of gamma radiation on in vitro shoot forming capacity of cotyledon explants in red pepper (Capsicum annuum L. cv. Yatsufusa). Jap. J. Breed. 38, 27–34.
Su Y.H., Liu Y.B., Zhang X.S., 2011. Auxin-cytokinin interaction regulates meristem development. Mol. Plant 4(4), 616–625.
Szasz A., Nervo G., Fari M., 1995. Screening for in vitro shoot-forming capacity of seedling explants in bell pepper (Capsicum annuum L.) genotypes and efficient plant regeneration using thidiazuron. Plant Cell. Rep. 14, 666–669.
Andrzej Gatz
University of Technology and Life Sciences in BydgoszczUniversity of Technology and Life Sciences in Bydgoszcz
University of Technology and Life Sciences in BydgoszczUniversity of Technology and Life Sciences in Bydgoszcz
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