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In vitro selection and characterization of buffelgrass somaclones with different responses to water stress

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Abstract

Buffelgrass is a forage grass that reproduces mainly by apomixis. In species with this reproduction mode, in vitro selection allows the incorporation of alternatives in a breeding program. The aims were to define a protocol for in vitro selection, provide a molecular and morphological characterization of the progenies of regenerated plants, and evaluate them under water stress conditions. In the embryogenic callus induction medium (IM), the highest values of the variables fresh weight of embryogenic calli, proportion of embryogenic calli and number of regenerated seedlings (NRS) were obtained in the 25 mM mannitol treatment. The remaining concentrations of the osmotic agent (50, 75, 100 and 150 mM) had a negative effect on these variables. In the regeneration medium (RM), NRS was reduced at all mannitol concentrations. When embryogenic calli were induced and seedlings were regenerated maintaining mannitol concentrations in IM and RM, the highest NRS values were recorded at 25 mM mannitol. In vitro regenerated seedlings transplanted to an experimental plot exhibited different morphological characteristics from those of the anther donor plant. ISSR primers detected 22% of polymorphic bands and divergence between 0.20 and 0.37 in in vitro regenerated plants. Finally, water stress assays confirmed that S1 progenies exhibited a differential behavior from that of the parent material. Under 100 mM of mannitol used as selection pressure in IM or in both IM and RM, S1 progenies of two regenerated materials had higher height, fresh weight and dry weight at the end of water stress assay.

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Abbreviations

IM:

Induction medium

RM:

Regeneration medium

FWEC:

Fresh weight of embryogenic calli

PEC:

Proportion of embryogenic calli

NRS:

Number of regenerated seedlings

IS:

Induction stage

RS:

Regeneration stage

R1 :

Regenerated seedlings

S1 :

Progenies of in vitro regenerated plants

ISSR:

Inter simple sequence repeat

References

  • Aazami MA, Torabi M, Jalili E (2010) In vitro response of promising tomato genotypes for tolerance to osmotic stress. Afr J Biotechnol 9:4014–4017.

    CAS  Google Scholar 

  • Ahmad MSA, Javed F, Ashraf M (2007) Iso-osmotic effect of NaCl and PEG on growth, cations and free proline accumulation in callus tissue of two indica rice (Oryza sativa L.) genotypes. Plant Growth Regul 53:53–63

    Article  Google Scholar 

  • Aoshima Y (2005) Efficient embryogenesis in the callus of tea (Camellia sinensis) enhanced by the osmotic stress or antibiotics treatment. Plant Biotechnol 22:277–280

    Article  CAS  Google Scholar 

  • Bairu MW, Aremu AO, Van Staden J (2011) Somaclonal variation in plants: causes and detection methods. Plant Growth Regul 63:147–173

    Article  CAS  Google Scholar 

  • Balzarini MG, Di Rienzo JA (2013) InfoGen versión 2013. FCA, Universidad Nacional de Córdoba, Argentina. http://www.info-gen.com.ar

  • Bashaw EC, Hignight KW (1990) Gene transfer in apomictic buffelgrass through fertilization of an unreduced egg. Crop Sci 30:571–575

    Article  Google Scholar 

  • Bhat V, Dalton SJ, Kumar S, Bhat BV, Gupta MG, Morris P (2001) Particle-inflow-gun-mediated genetic transformation of buffelgrass (Cenchrus ciliaris L.): optimizing biological and physical parameters. J Appl Genet 42:405–412

    CAS  PubMed  Google Scholar 

  • Biahoua A, Bonneau L (1999) Control of in vitro somatic embryogenesis of the spindle tree (Euonymus europaeus L.) by the sugar type and the osmotic potential of the culture medium. Plant Cell Rep 19:185–190

    Article  CAS  Google Scholar 

  • Biswas J, Chowdhury B, Bhattacharya A, Mandal AB (2002) In vitro screening for increased drought tolerance in rice. In Vitro Cell Dev Plant 38:525–530

    Article  Google Scholar 

  • Carloni E (2016) Cultivo in vitro de anteras como estrategia para el mejoramiento genético de buffelgrass (Cenchrus ciliaris L). Tesis Doctorado. FCA – Universidad Nacional de Córdoba. https://rdu.unc.edu.ar/bitstream/handle/11086/2729/Carloni.%20Cultivo%20in%20vitro%20de%20anteras%20como%20estrategia%20para%20el%20mejoramiento%20gen%C3%A9tico%20de%20buffelgrass...%20.pdf?sequence=1

  • Carloni E, Ribotta A, Colomba EL, Griffa S, Quiroga M, Tommasino E, Grunberg K (2014) Somatic embryogenesis from in vitro anther culture of apomictic buffel grass genotypes and analysis of regenerated plants using flow cytometry. Plant Cell Tissue Organ Cult 117:311–322

    Article  Google Scholar 

  • Cha-um S, Takabe T, Kirdmanee C (2012) Physio-biochemical responses of oil palm (Elaeis guineensis Jacq.) seedlings to mannitol and polyethylene glycol-induced iso-osmotic stresses. Plant Prod Sci 15:65–72

    Article  CAS  Google Scholar 

  • Claeys H, Van Landeghem S, Dubois M, Maleux K, Inzé D (2014) What is stress? Dose-response effects in commonly used in vitro stress assays. Plant Physiol 165:519–527

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Colomba EL, Grunberg K, Griffa S, Ribotta A, Mroginski L, Biderbost E (2006) The effect of genotype and culture medium on somatic embryogenesis and plant regeneration from mature embryos of fourteen apomictic cultivars of buffelgrass (Cenchrus ciliaris L.). Grass Forage Sci 61:2–8

    Article  Google Scholar 

  • Conti H, Cazenave G, Giagnoni R (2009) Características climáticas de las provincias de Santiago del Estero, Chaco y Formosa. Flora Chaqueña, Asteraceae. Colecc Ci Inst Tecnol Agropecu 23:9–26

    Google Scholar 

  • Di Rienzo JA, Guzmán AW, Casanoves F (2002) A multiple comparisons method based on the distribution of the root node distance of a binary tree. J Agric Biol Environ Stat 7:1–14

    Article  Google Scholar 

  • Di Rienzo JA, Casanoves F, Balzarini MG, Gonzalez L, Tablada M, Robledo CW (2014) Grupo InfoStat, FCA, Universidad Nacional de Córdoba, Argentina

  • Errabii T, Gandonou CB, Essalmani H, Abrini J, Idaomar M, Senhaji NS (2007) Effects of NaCl and mannitol induced stress on sugarcane (Saccharum sp.) callus cultures. Acta Physiol Plant 29:95–102

    Article  CAS  Google Scholar 

  • Gangopadhyay G, Basu S, Gupta S (1997) In vitro selection and physiological characterization of NaCl-and mannitol-adapted callus lines in Brassica juncea. Plant Cell Tissue Organ Cult 50:161–169

    Article  CAS  Google Scholar 

  • Griffa S, Ribotta A, López Colomba E, Tommasino E, Carloni E, Luna C, Grunberg K (2010) Evaluation seedling biomass and its components as selection criteria for improving salt tolerance in Buffel grass genotypes. Grass Forage Sci 65:358–361

    Article  Google Scholar 

  • Gulati A, Jaiwal PK (1993) Selection and characterization of mannitol-tolerant callus lines of Vigna radiata (L.) Wilczek. Plant Cell Tissue Organ Cult 34:35–41

    Article  CAS  Google Scholar 

  • Gupta M, Chyi YS, Romero-Severson J, Owen JL (1994) Amplification of DNA markers from evolutionarily diverse genomes using single primers of simple-sequence repeats. Theor Appl Genet 89:998–1006

    CAS  PubMed  Google Scholar 

  • Hanna WW, Bashaw EC (1987) Apomixis: its identification and use in plant breeding. Crop Sci 27:1136–1139

    Article  Google Scholar 

  • Hanselka CW, Hussey MA, Ibarra F (2004) Buffelgrass. In: Moser LE, Burson BL, Sollenberger LE (eds) Warm-season (C4) grasses. American Society of Agronomy, Madison, pp 477–502

    Google Scholar 

  • Hassan NS, Shaaban LD, Hashem ESA, Seleem EE (2004) In vitro selection for water stress tolerant callus line of Helianthus annus L. Cv. Myak. Int J Agric Biol 6:1–13

    Google Scholar 

  • Hilae A, Te-chato S (2005) Effect of carbon sources and strength of MS medium on germination of somatic embryos of oil palm (Elaeis guineensis Jacq.). Songklanakarin J Sci Technol 27:629–635.

    Google Scholar 

  • Hohl M, Schopfer P (1991) Water relations of growing Maize coleoptiles comparison between mannitol and polyethylene glycol 6000 as external osmotica for adjusting turgor pressure. Plant Physiol 95:716–722

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jessup RW, Burson BL, Burow O, Wang YW, Chang C, Li Z, Paterson AH, Hussey MA (2003) Segmental allotetraploidy and allelic interactions in buffelgrass (Pennisetum ciliare (L.) Link syn. Cenchrus ciliaris L.) as revealed by genome mapping. Genome 46:304–313

    Article  CAS  PubMed  Google Scholar 

  • Joshi R, Shukla A, Sairam RK (2011) In vitro screening of rice genotypes for drought tolerance using polyethylene glycol. Acta Physiol Plant 33:2209–2217

    Article  CAS  Google Scholar 

  • Kamada H, Ishikawa K, Saga H, Harada H (1993) Induction of somatic embryogenesis in carrot by osmotic stress. Plant Tissue Cult Lett 10:38–44

    Article  CAS  Google Scholar 

  • Karami O, Saidi A (2010) The molecular basis for stress-induced acquisition of somatic embryogenesis. Mol Biol Rep 37:2493–2507

    Article  CAS  PubMed  Google Scholar 

  • Karp A (1995) Somaclonal variation as a tool for crop improvement. Euphytica 85:295–302

    Article  Google Scholar 

  • Kumar S, Bhat V (2012) High-frequency direct plant regeneration via multiple shoot induction in the apomictic forage grass Cenchrus ciliaris L. In Vitro Cell Dev Plant 48:241–248

    Article  Google Scholar 

  • Kumar S, Sahu N, Singh A (2015) High-frequency in vitro plant regeneration via callus induction in a rare sexual plant of Cenchrus ciliaris L. In Vitro Cell Dev Plant 51:28–34

    Article  CAS  Google Scholar 

  • Larkin PJ, Scowcroft SC (1981) Somaclonal variation: a novel source of variability from cell culture for plant improvement. Theor Appl Genet 60:197–214

    Article  CAS  PubMed  Google Scholar 

  • Leone A, Costa A, Tucci M, Grillo S (1994) Adaptation versus shock response to polyethylene glycol-induced low water potential in cultured potato cells. Physiol Plant 92:21–30

    Article  CAS  Google Scholar 

  • Lipavská H, Vreugdenhil D (1996) Uptake of mannitol from the media by in vitro grown plants. Plant Cell Tissue Organ Cult 45:103–107

    Article  Google Scholar 

  • López Colomba E (2011) Inducción de variabilidad genética para tolerancia a estreses abióticos mediante técnicas de cultivo in vitro en Cenchrus ciliaris L. Tesis Maestría. Universidad Internacional de Andalucía. http://dspace.unia.es/bitstream/handle/10334/1681/0229_Lopez.pdf?sequence=1

  • López Colomba E, Prina A, Griffa S, Ribotta AN, Carloni E, Tommasino E, Luna C, Biderbost E, Grunberg K (2011). Obtaining new germplasm in Cenchrus ciliaris L. through induced-mutation and in vitro selection. Phyton Int J Exp Bot 80:59–64

    Google Scholar 

  • López Colomba E, Tommasino E, Luna C, Griffa S, Carloni E, Ribotta A, Quiroga M, Grunberg K (2013) Differential salt-stress response during germination and vegetative growth in in vitro selected somaclonal mutants of Cenchrus ciliaris L. S Afr J Bot 87:157–163

    Article  Google Scholar 

  • Lu S, Peng X, Guo Z, Zhang G, Wang Z, Wang C, Pang C, Fan Z, Wang J (2007) In vitro selection of salinity tolerant variants from triploid bermudagrass (Cynodon transvaalensis × C. dactylon) and their physiological responses to salt and drought stress. Plant Cell Rep 26:1413–1420

    Article  CAS  PubMed  Google Scholar 

  • Lu S, Chen C, Wang Z, Guo Z, Li H (2009) Physiological responses of somaclonal variants of triploid bermudagrass (Cynodon transvaalensis × Cynodon dactylon) to drought stress. Plant Cell Rep 28:517–526

    Article  CAS  PubMed  Google Scholar 

  • Mahmood I, Razzaq A, Hafiz IA, Kaleem S, Khan AA, Qayyum A, Ahmad M (2012) Interaction of callus selection media and stress duration for in vitro selection of drought tolerant callus of wheat. Afr J Biotechnol 11:4000–4006

    CAS  Google Scholar 

  • Matheka JM, Magiri E, Rasha AO, Machuka J (2008) In vitro selection and characterization of drought tolerant somaclones of tropical maize (Zea mays L.). Biotechnology 7:641–650

    Article  CAS  Google Scholar 

  • Misra S, Attree SM, Leal I, Fowke LC (1993) Effect of abscisic acid, osmoticum, and dessication on synthesis of storage proteins during the development of white spruce somatic embryos. Ann Bot 71:11–22

    Article  CAS  Google Scholar 

  • Mohamed MAH, Ibrahim TA (2012) Enhanced in vitro production of Ruta graveolens L. coumarins and rutin by mannitol and ventilation. Plant Cell Tissue Organ Cult 111:335–343

    Article  CAS  Google Scholar 

  • Mohamed MH, Harris PJC, Henderson J (2000) In vitro selection and characterisation of a drought tolerant clone of Tagetes minuta. Plant Sci 159:213–222

    Article  CAS  PubMed  Google Scholar 

  • Moore KJ, Boote KJ, Sanderson MA (2004) Physiology and developmental morphology. In: Moser LE, Burson BL, Sollenberger LE (eds) Warm-season (C4) grasses. American Society of Agronomy, Madison, pp 179–216

    Google Scholar 

  • Munns R (2002) Comparative physiology of salt and water stress. Plant Cell Environ 25:239–250

    Article  CAS  PubMed  Google Scholar 

  • Nadel BL, Altman A, Ziv M (1989) Regulation of somatic embryogenesis in celery cell suspensions. Plant Cell Tissue Organ Cult 18:181–189

    Article  CAS  Google Scholar 

  • Pérez H (2005) Características de las especies forrajeras adaptadas a las condiciones del Noroeste del país. Forrajes 33–41. http://www.produccionanimal.com.ar/produccion_y_manejo_pasturas/pasturas_cultivadas_megatermicas/68-caforra.pdf

  • Phillips RL, Kaeppler SM, Olhoft P (1994) Genetic instability of plant tissue cultures: breakdown of normal controls. Proc Natl Acad Sci USA 91:5222–5226

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Quiroga M, Grunberg K, Ribotta A, López Colomba E, Carloni E, Tommasino E, Luna C, Griffa S (2013) Obtaining sexual genotypes for breeding in buffel grass. S Afr J Bot 88:118–123

    Article  Google Scholar 

  • Rejšková A, Patková L, Stodůlková E, Lipavská H (2007) The effect of abiotic stresses on carbohydrate status of olive shoots (Olea europaea L.) under in vitro conditions. J Plant Physiol 164:174–184

    Article  PubMed  Google Scholar 

  • Remotti PC (1998) Somaclonal variation and in-vitro selection for crop improvement. In: Jain SM, Brar DS, Ahloowali BS (eds) Somaclonal variation and induced mutations in crop improvement. Kluwer Academic Publishers, Dordrecht, pp 169–201

    Chapter  Google Scholar 

  • Stasolla C, van Zyl L, Egertsdotter U, Craig D, Liu W, Sederoff RR (2003) The effects of polyethylene glycol on gene expression of developing white spruce somatic embryos. Plant Physiol 131:49–60

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Svobodová H, Albrechtová J, Kumstýřová L, Lipavská H, Vágner M, Vondráková Z (1999) Somatic embryogenesis in Norway spruce: anatomical study of embryo development and influence of polyethylene glycol on maturation process. Plant Physiol Biochem 37:209–221

    Article  Google Scholar 

  • Tommasino E (2013) I Workshop sobre Estresses Abióticos em Plantas Forrageiras. Disertación: Fenotipificación de Genotipos de Cenchrus ciliaris para tolerancia al estrés hídrico. http://cloud.cnpgc.embrapa.br/weapf2013/files/2012/12/ENSAYOS-DE-ESTR%C3%89S-POR-SEQU%C3%8DA-FINAL.pdf

  • Tommasino E, Griffa S, Grunberg K, Ribotta A, López Colomba E, Carloni E, Quiroga M, Luna C (2012) Malondialdehyde content as a potential biochemical indicator of tolerant Cenchrus ciliaris L. genotypes under heat stress treatment. Grass Forage Sci 67:456–459

    Article  CAS  Google Scholar 

  • Verma D, Ansari MW, Agrawal GK, Rakwal R, Shukla A, Tuteja N (2013) In vitro selection and field responses of somaclonal variant plants of rice cv PS113 for drought tolerance. Plant Signal Behav 8:23519

    Article  Google Scholar 

  • Verslues PE, Ober ES, Sharp RE (1998) Root growth and oxygen relations at low water potentials. Impact of oxygen availability in polyethylene glycol solutions. Plant Physiol 116:1403–1412

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Verslues PE, Agarwal M, Katiyar Agarwal S, Zhu J, Zhu JK (2006) Methods and concepts in quantifying resistance to drought, salt and freezing, abiotic stresses that affect plant water status. Plant J 45:523–539

    Article  CAS  PubMed  Google Scholar 

  • Vítová L, Stodůlková E, Bartoníčková A, Lipavská H (2002) Mannitol utilisation by celery (Apium graveolens) plants grown under different conditions in vitro. Plant Sci 163:907–916

    Article  Google Scholar 

  • Yaseen M, Ahmad T, Abbasi NA, Hafiz IA (2009) Assessment of apple rootstocks M 9 and M 26 for in vitro rooting potential using different carbon sources. Pak J Bot 41:769–781

    Google Scholar 

  • Yaseen M, Ahmad T, Sablok G, Standardi A, Hafiz IA (2013) Review: role of carbon sources for in vitro plant growth and development. Mol Biol Rep 40:2837–2849

    Article  CAS  PubMed  Google Scholar 

  • Zavattieri MA, Frederico AM, Lima M, Sabino R, Arnholdt-Schmitt B (2010) Induction of somatic embryogenesis as an example of stress-related plant reactions. Electron J Biotechnol 13:1–9

    Article  Google Scholar 

  • Zietkiewicz E, Rafalski A, Labuda D (1994) Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20:176–183

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported financially by The Instituto Nacional de Tecnología Agropecuaria (INTA), Argentina, Project PNPA-1126072 and CONICET PIP 112 201101 0031.

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Authors and Affiliations

  1. Instituto de Fisiología y Recursos Genéticos Vegetales (IFRGV), Centro de Investigaciones Agropecuarias (CIAP), Instituto Nacional de Tecnología Agropecuaria (INTA), Av. 11 de Septiembre 4755, X5020ICA, Córdoba, Argentina

    Edgardo Carloni, Exequiel Tommasino, Eliana López Colomba, Andrea Ribotta, Mariana Quiroga, Sabrina Griffa & Karina Grunberg

  2. Consejo de Nacional Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, C1425FQB, CABA, Argentina

    Karina Grunberg

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  1. Edgardo Carloni
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  2. Exequiel Tommasino
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  3. Eliana López Colomba
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  4. Andrea Ribotta
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  5. Mariana Quiroga
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Correspondence to Edgardo Carloni.

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Carloni, E., Tommasino, E., López Colomba, E. et al. In vitro selection and characterization of buffelgrass somaclones with different responses to water stress. Plant Cell Tiss Organ Cult 130, 265–277 (2017). https://doi.org/10.1007/s11240-017-1220-9

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