[en] An efficacious regeneration system was developed from shoot apex explants of Zea mays using a two-step culture procedure. Seventeen Indian genotypes were assessed for their regeneration potential. The maximum response of shoot induction was obtained from explants cultured on Murashige and Skoog medium supplemented with 4.5 µM thidiazuron and 26.7 µM glycine. Maximum mean number of shoots (17.2) was observed in genotype COH (m)-5 while NPK was the least responsive (6.7). Shoot clumps transferred from shoot induction medium to multiplication (second) medium amended with 1.1 µM thidiazuron and 0.88 µM N 6 -benzylaminopurine showed increased number of shoots in COH (m)-5 (36.1 shoots); NPK was the least responsive with an average of 9.5 shoots. The best response in root induction, with a larger number of roots (10.5) and longer roots (6.6 cm), was observed in Murashige and Skoog medium supplemented with 7.3 µM indole-3-butyric acid and 7.9 µM phloroglucinol. Analysis of variance indicated that plant regeneration response varied greatly among the genotypes. In vitro raised plants were successfully transferred to the field after hardening, with a 99 % survival rate. Inter simple sequence repeats analysis revealed that the similarity matrix pair-wise value was 1, the Mantel test value was p 1.0; Analysis of molecular variance genetic variances were 93 % within the population and 7 % between populations; Principal component jolliffe cut off was 0.15, Principal component and Principle coordinate analysis % variance was 13.19. These values were congruent for both the mother and the in vitro-raised plants, confirming genetic integrity.
Disciplines :
Biotechnology
Author, co-author :
Ramakrishnan, M.
Stanislaus, Antony Ceasar ; Université de Liège - ULiège > Département des sciences de la vie > Génomique fonctionnelle et imagerie moléculaire végétale
Duraipandiyan, V
Ignacimuthu, S
Language :
English
Title :
Efficient plant regeneration from shoot apex explants of maize (Zea mays) and analysis of genetic fidelity of regenerated plants by ISSR markers
Aggarwal D, Kumar A, Reddy MS (2010) Shoot organogenesis in elite clones of Eucalyptus tereticornis. Plant Cell Tiss Organ Cult 102:45–52
Aguado-Santacruz GA, Garcia-Moya E, Aguilar-Acuna JL, Moreno-Gomez B, SolisMoya E, Preciado-Ortiz ER, Jimenez-Bremont JF, Rascon-Cruz Q (2007) In vitro plant regeneration from quality protein maize (QPM). In Vitro Cell Dev Biol Plant 43:215–224
Ahmadabadi M, Ruf S, Bock R (2007) A leaf based regeneration and transformation system for maize (Zea mays L.). Transgenic Res 16:437–448
Al-Abed D, Rudrabhatla S, Talla R, Goldman S (2006) Split seed: a new tool for maize researchers. Planta 223:1355–1360
Anthony JH (1986) The maximum familywise error rate of Fisher’s least significant difference test. J Am Stat Assoc 81:1000–1004
Arshad M, Silvestre J, Merlina G, Dumat C, Pinelli E, Kallerhoff J (2012) Thidiazuron-induced shoot organogenesis from mature leaf explants of scented Pelargonium capitatum cultivars. Plant Cell Tiss Organ Cult 108(2):315–322
Bakshi S, Roy NK, Sahoo L (2012) Seedling preconditioning in thidiazuron enhances axillary shoot proliferation and recovery of transgenic cowpea plants. Plant Cell Tiss Organ Cult 110:77–91
Barloy D, Beckert M (1993) Improvement of regeneration ability of androgenetic embryos by early anther transfer in maize plant. Plant Cell Tiss Organ Cult 33:45–50
Bhatia R, Singh KP, Sharma TR, Jhang T (2011) Evaluation of the genetic fidelity of in vitro-propagated gerbera (Gerbera jamesonii Bolus) using DNA-based markers. Plant Cell Tiss Organ Cult 104:131–135
Carra A, Sajeva M, Abbate L, Siragusa M, Sottile F, Carimi F (2012) In vitro plant regeneration of caper (Capparis spinosa L.) from floral explants and genetic stability of regenerants. Plant Cell Tiss Organ Cult 109:373–381
Ceasar SA, Ignacimuthu S (2008) Efficient somatic embryogenesis and plant regeneration from shoot apex explants of different Indian genotypes of finger millet (Eleusine coracana (L.) Gaertn.). In Vitro Cell Dev Biol Plant 44:427–435
Ceasar SA, Ignacimuthu S (2010) Effects of cytokinins, carbohydrates and amino acids on induction and maturation of somatic embryos in kodo millet (Paspalum scorbiculatum Linn.). Plant Cell Tiss Organ Cult 102:153–162
Ceasar SA, Lenin Maxwell S, Bhargav Prasad K, Karthigan M, Ignacimuthu S (2010) Highly efficient shoot regeneration of Bacopa monnieri (L.) using a two-stage culture procedure and assessment of genetic integrity of micropropagated plants by RAPD. Acta Physiologiae Plantarum 32(3):443–452
Chen F, Shi J, Luo Y-q, Pu M, Sun S-y (2013) Genetic characterization of the Gypsy Moth from China (Lepidoptera, Lymantriidae) using inter simple sequence repeats markers. PLoS ONE 8(8):e73017
Dai W, Su Y, Castillo C, Beslot O (2010) Plant regeneration from in vitro leaf tissues of Viburnum dentatum L. Plant Cell Tiss Organ Cult 104:257–262
De Klerk GJ, Vander Kreiken W, De Jong JC (1999) The formation of adventitious roots: new concepts, new possibilities. In Vitro Cell Dev Biol Plant 35:189–199
Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15
Filippov M, Miroshnichenko D, Vernikovskaya D (2006) Sergey the effect of auxins, time exposure to auxin and genotypes on somatic embryogenesis from mature embryos of wheat. Plant Cell Tiss Organ Cult 84:100192–100201
Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:51–158
Giridhar P, Gururaj H, Ravishankar G (2005) In vitro shoot multiplication through shoot tip cultures of Decalepis hamiltonii Wight & Arn., a threatened plant endemic to Southern India. In Vitro Cell Dev Biol Plant 41:77–80
Green CE, Philips RL (1975) Plant regeneration from tissue culture of maize. Crop Sci 15:417–421
Gupta PK, Roy JK (2002) Molecular markers in crop improvement: present status and future needs in India. Plant Cell Tiss Organ Cult 70:229–234
Hammer Ø, Harper DAT, Ryan PD (2001) PAST: paleontological statistics software package for education and data analysis. Palaeontologia Electronica 4(1):9
Huang D, Dai W (2011) Direct regeneration from in vitro leaf and petiole tissues of Populus tremula ‘Erecta’. Plant Cell Tiss Organ Cult 107:169–174
Huang XQ, Wei ZM (2004) High frequency plant regeneration through callus initiation from mature embryos of maize. Plant Cell Rep 22:793–800
Huetteman CA, Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tiss Organ Cult 33:105–119
Jaccard P (1908) Nouvelles researches sur la distribution florale. Bull Soc vaudoise Sci Natl 44:223–270
Jolliffe IT (2002) Principal component analysis, 2nd edn. Springer, New York
Joshi P, Dhawan V (2007) Assesment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assey. Biol Plant 51:22–26
Kishor R, Devi HS (2009) Induction of multiple shoots in a monopodial orchid hybrid (Aerides vandarum Reichb. f 3 Vanda stangeana Reichb.f) using thidiazuron and analysis of their genetic stability. Plant Cell Tiss Organ Cult 97:121–129
Kou Y, Ma G, Da Silva JA, Liu N (2013) Callus induction and shoot organogenesis from anther cultures of Curcuma attenuata Wall. Plant Cell Tiss Organ Cult 112(1):1–7
Kwapata K, Sabzikar R, Sticklen MB, Kelly JD (2010) In vitro regeneration and morphogenesis studies in common bean. Plant Cell Tiss Organ Cult 100:97–105
Leory XJ, Leon K, Hily JM, Chaumeli P, Branchard M (2001) Detection of in vitro culture-induced instability through intersimple sequence repeat analysis. Theor Appl Genet 102:885–891
Levi M, Thomas CE, Newman M, Reddy OUK, Zhang X, Xu Y (2004) ISSR and AFLP markers differ among American watermelon cultivars with limited genetic diversity. J Am Soc Hortic Sci 129(4):553–558
Li W, Sun G, Liu J, Masilamany P, Taylor JH, Yan W, Kasha KJ, Paul KP (2004) Inheritance of plant regeneration from maize (Zea mays L.) shoot meristem cultures derived from germinated seeds and the identification of associated RAPD and SSR markers. Theor Appl Genet 108:681–687
Li Z, Mize K, Campbell F (2010) Regeneration of daylily (Hemerocallis) from young leaf segments. Plant Cell Tiss Organ Cult 102:199–204
Lin GZ, Zhao XM, Hong SK, Lian YJ (2011) Somatic embryogenesis and shoot organogenesis in the medicinal plant Pulsatilla koreana Nakai. Plant Cell Tiss Organ Cult 106:93–103
Liu F, Huang Li-Li, Li Y-Li, Reinhoud P, Jongsma MA, Wang C-Y (2011) Shoot organogenesis in leaf explants of Hydrangea macrophylla ‘Hyd1’ and assessing genetic stability of regenerants using ISSR markers. Plant Cell Tiss Organ Cult 104:111–117
Lowe K, Bowen B, Hoerster G, Ross M, Bond D, Pierce D, Gordon-Kamm B (1995) Germline transformation of maize following manipulation of chimeric shoot meristems. Nat Biotechnol 13:677–682
Ma G, da Silva JAT, Lu J, Zhang X, Zhao J (2010) Shoot organogenesis and plant regeneration in Metabriggsia ovalifolia. Plant Cell Tiss Organ Cult 105(3):355–361
Magyar-Tabori K, Dobranszki J, Da Silva JAT, Bulley SM, Hudak I (2010) The role of cytokinins in shoot organogenesis in apple. Plant Cell Tiss Organ Cult 101(3):251–267
Manivannan A, Kaul J, Singode A, Dass S (2010) Callus induction and regeneration of elite Indian maize inbreds. Afr J Biotechnol 9(44):7446–7452
Mishra J, Singh M, Palni LMS, Nandi SK (2011) Assessment of genetic fidelity of encapsulated microshoots of Picrorhiza kurrooa. Plant Cell Tiss Organ Cult 104:181–186
Mithila J, Murch S, KrishnaRaj S, Saxena PK (2001) Recent advances in Pelargonium in vitro regeneration systems. Plant Cell Tiss Organ Cult 67:1–9
Mohanty S, Parida R, Singh S, Joshi RK, Subudhi E, Nayak S (2011) Biochemical and molecular profiling of micropropagated and conventionally grown Kaempferia galangal. Plant Cell Tiss Organ Cult 106:39–46
Morocz C, Donn G, Nemeth J, Dudits D (1990) An improved system to obtain fertile regenerants via maize protoplast isolated from highly embryogenic suspension culture. Theor Appl Genet 80:721–726
Muoma J, Muluvi G, Machuka J (2008) In vitro regeneration by indirect organogenesis of selected Kenyan maize genotypes using shoot apex. Biotechnology 7(4):732–738
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Mythili P, Satyavati V, Pavankumar G, Rao MV, Manga V (1997) Genetic analysis of short term callus culture and orphogenesis in pearl millet, Pennisetum glacum. Plant Cell Tiss Organ Cult 50:171–178
Nitnaware KM, Naik DG, Nikam TD (2010) Thidiazuron-induced shoot organogenesis and production of hepatoprotective lignan phyllanthin and hypophyllanthin in Phyllanthus amarus. Plant Cell Tiss Organ Cult 104(1):101–110
O’Connor-Sanchez A, Cabrera-Ponce JL, Valdez-Melara M, Tellez-Rodriguez P, Pons-Hernandez JL, Herrera-Estrella L (2002) Transgenic maize plants of tropical and subtropical genotypes obtained from calluses containing organogenic and embryogenic-like structures derived from shoot tips. Plant Cell Rep 21:302–312
Pareddy DR, Petolino JF (1990) Somatic embryogenesis and plant regeneration from immature inflorescences of several elite inbreds of maize. Plant Sci 46:225–232
Pathi KM, Tula S, Huda KMK, Srivastava VK, Tuteja N (2013) An efficient and rapid regeneration via multiple shoot induction from mature seed derived embryogenic and organogenic callus of Indian maize (Zea mays L.). Plant Signal Behav 8(10):1–6
Peakall R, Smouse PE (2012) GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research-an update. Bioinformatics 28(19):2537–2539
Pinheiro LR, Rabbani ARC, da Silva AVC, da Ledo Silva A, Pereira KLG, Diniz LEC (2012) Genetic diversity and population structure in the Brazilian Cattleya labiata (Orchidaceae) using RAPD and ISSR markers. Plant Syst Evol 298(10):1815–1825
Rafiq M, Fatima T, Husnain T, Bashir K, Riazuddin S (2005) Effect of different media on callus formation and regeneration of different genotypes of Maize. Plant Tissue Cult 15(1):57–65
Rahman M, Rajora O (2001) Microsatellite DNA somaclonal variation in micropropagated trembling aspen (Populus tremuloides). Plant Cel Rep 20:531–536
Rai MK, Phulwaria M, Harish, Gupta AK, Shekhawat NS, Jaiswal U (2012) Genetic homogeneity of guava plants derived from somatic embryogenesis using SSR and ISSR markers. Plant Cell Tiss Organ Cult 111:259–264
Rakshit S, Rashid Z, Sekhar JC, Fatma T, Dass S (2010) Callus induction and whole plant regeneration in elite Indian maize (Zea mays L.) inbreds. Plant Cell Tiss Organ Cult 100:31–37
Ramakrishnan M, Ceasar SA, Duraipandiyan V, Melvin AD, Ignacimuthu S (2013) Efficacious somatic embryogenesis and fertile plant recovery from shoot apex explants of onion (Allium cepa. L.). In Vitro Cell Dev Biol Plant 49:285–293
Ray T, Dutta I, Saha P, Das S, Roy SC (2006) Genetic stability of three economically important micropropagated banana (Musa spp.) cultivars of lower Indo-Gangetic plains, as assessed by RAPD and ISSR markers. Plant Cell Tiss Organ Cult 85:11–21
Reddy MP, Sarla N, Siddiq EA (2002) Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica 128:9–17
Sairam RV, Paran M, Franklin G, Lifeng Z, Smith B, MacDougall J, Wilber C, Sheikhi H, Kashikar N, Meeker K, Al-Abed D, Berry K, Vierling R, Goldman SL (2003) Shoot meristem an ideal explant for Zea mays L. transformation. Genome 46:323–329
Sarkar D, Naik PS (2000) Phloroglucinol enhances growth and rate of axillary shoot proliferation in potato shoot tip culture in vitro. Plant Cell Tiss Organ Cult 60:139–149
Semagn K, Bjørnstad A, Ndjiondjop MN (2006) An overview of molecular marker methods for plants. Afr J Biotechnol 5(25):2540–2568
Shaik S, Singh N, Nicholas A (2011) Cytokinin-induced organogenesis in Lessertia (Sutherlandia) frutescens L. using hypocotyl and cotyledon explants affects yields of L-canavanine in shoots. Plant Cell Tiss Organ Cult 105:439–446
Shilpha J, Silambarasan T, Largia MJV, Ramesh M (2014) Improved in vitro propagation, solasodine accumulation and assessment of clonal fidelity in regenerants of Solanum trilobatum L. by flow cytometry and SPAR methods. Plant Cell Tiss Organ Cult 117:125–129
Songstad DD, Peterson WL, Armstrong CL (1992) Establishment of friable embryogenic (type II) callus from immature tassels of Zea mays (Poaceae). Am J Bot 79:761–764
SPSS Inc (2003) SPSS base 12.0 user’s guide. SPSS Inc, Chicago
Sriskandarajah S, Lundquist PO (2009) High frequency shoot organogenesis and somatic embryogenesis in juvenile and adult tissues of seabuckthorn (Hippophae rhamnoides L.). Plant Cell Tiss Organ Cult 99(3):259–268
Sujatha M, Vijay S, Vasavi S, Sivaraj N, Rao SC (2012a) Combination of thidiazuron and 2-isopentenyladenine promotes highly efficient adventitious shoot regeneration from cotyledons of mature sunflower seeds (Helianthus annuus L.). Plant Cell Tiss Organ Cult 111(3):359–372
Sujatha M, Vijay S, Vasavi S, Veera Reddy P, Chander Rao S (2012b) Agrobacterium-mediated transformation of cotyledons of mature seeds of multiple genotypes of sunflower (Helianthus annuus L.). Plant Cell Tiss Organ Cult 110:275–287
Tiwari V, Tewari KN, Singh BD (2001) Comparative studies of cytokinin on in vitro propagation of Bacopa monniera. Plant Cell Tiss Organ Cult 66:9–16
Verma P, Mathur AK (2011) Direct shoot bud organogenesis and plant regeneration from pre-plasmolysed leaf explants in Catharanthus roseus. Plant Cell Tiss Organ Cult 106:401–408
Vladimir S, Gilbertson L, Adae P, Duncan D (2006) Agrobacterium-mediated transformation of seedling-derived maize callus. Plant Cell Rep 25:320–328
Wang QM, Gao FZ, Gao X, Zou FY, Sui X, Wang M, Hui YJ, Wang L (2011) Regeneration of Clivia miniata and assessment of clonal fidelity of plantlets. Plant Cell Tiss Organ Cult 109(2):191–200
Winkelmann T, Kaviani K, Serek M (2005) Development of a shoot regeneration protocol for genetic transformation in Pelargonium zonale and Pelargonium peltatum hybrids. Plant Cell Tiss Organ Cult 80:33–42
Yang X, Lu J, Da Silva JA, Ma G (2012) Somatic embryogenesis and shoot organogenesis from leaf explants of Primulina tabacum. Plant Cell Tiss Organ Cult 109:213–221
Yang L, Bian L, Shen HL, Li YH (2013) Somatic embryogenesis and plantlet regeneration from mature zygotic embryos of Manchurian ash (Fraxinus mandshurica Rupr.). Plant Cell Tiss Organ Cult 115:115–125
Zale JM, Borchand H, Kidwell KK, Stebber CM (2004) Callus induction and plant regeneration from mature embryo of diverse set of wheat genotypes. Plant Cell Tiss Organ Cult 76:277–281
Zanol GC, Fortes GR, De L, Campos AD, Centellas AQ, Da Silva JB (1998) In vitro rooting and peroxidase activity of apple rootstock treated with indole butyric acid and phloroglucinol. Res Brasil Fisiol Veg 10:65–68
Zhang S, Williams-Carrier R, Lemaux PG (2002) Transformation of recalcitrant maize elite inbreds using in vitro shoot meristematic cultures induced from germinated seedlings. Plant Cell Rep 21:263–270
Zhong H, Srinivasan C, Sticklen MB (1992) In-vitro morphogenesis of corn (Zeu mays L.). I. Differentiation of multiple shoot clumps and somatic embryos from shoot tips. Planta 187:483–489
Zhong H, Sun B, Warkentin D, Zhang S, Wu R, Wu T, Sticklen MB (1996) The competence of Maize shoot meristems for integrative transformation and inherited expression of transgenes. Plant Physiol 110:1097–1107