Biohardening of Arabidopsis thaliana Seeds and Seedlings with Fraser Photinia Associated Bacterium (PGB_invit) in In vitro Conditions


  • Maria BATOOL Gebze Technical University, Faculty of Basic Sciences, Department of Molecular Biology and Genetics, 41400, Kocaeli (TR)
  • Özlem AKKAYA Gebze Technical University, Faculty of Basic Sciences, Department of Molecular Biology and Genetics, 41400, Kocaeli (TR)
  • Mine GÜL ŞEKER Gebze Technical University, Faculty of Basic Sciences, Department of Molecular Biology and Genetics, 41400, Kocaeli (TR)
  • Yelda ÖZDEN ÇİFTÇİ Gebze Technical University, Faculty of Basic Sciences, Department of Molecular Biology and Genetics, 41400, Kocaeli (TR)



Arabidopsis thaliana, endophytic bacteria, plant-bacteria interactions, virD1 gene


The aim of this study was to analyze possible positive effects of putatively endophytic PGPB (PGB_invit), which was isolated from long-term in vitro cultured fraser photinia microshoots, on seed and 7-day old seedling stages of Arabidopsis thaliana. Seeds and in vitro-germinated seedlings were inoculated with 107 CFU/mL and 108 CFU/mL active (A) and inactive (I) endophytic bacterial populations along with their mix compositions (A+I) and suspended in MPYE broth together with their controls (untreated ones). 14 days old seedlings were evaluated for various plant growth parameters [i.e., shoot and root fresh weight (FW), shoot length (SL), shoot and root dry weight (DW), root length (RL) and photosynthetic pigments including chlorophyll a, b and carotenoids of plantlets] as well as endophytic and rhizospheric trait of bacteria. Positive effects of inactive and active bacterium on FW, DW and photosynthetic pigments for 7-day inoculated seedlings were recorded whereas an increase in photosynthetic contents for seed stage inoculations was observed. Rhizospheric and endophytic colonization of the bacteria was confirmed by PCR with the presence of virD1 gene, which is previously recorded to be existed in the plasmid of bacterium after inoculation. Overall, these results demonstrated that this peculiar putative endophytic bacterium being beneficial in active and even more useful in inactive form for A. thaliana when optimum conditions and concentrations are used. Moreover, presence of virD1 gene suggested its potential possibility to be used in bioengineering along with various other beneficial PGPR features as biofertilizer.


Bungard RA, Mcneil D, Morton J (1997). Effects of chilling, light and nitrogen-containing compounds on germinations, rate of germination and imbibition of Clematis vitalba L. Annals of Botany 79(6):643-645.

Dunne C, Crowley JJ, Loccoz YM, Dowling DN, de Bruijn F, O’Gara F (1993). Biological control of Pythium ultimum by Stenotrophomonas maltophilia W81 is mediated by an extracellular proteolytic activity. Microbiology 143(12):3921-3931.

Farh ME-A, Kim Y-J, Sukweenadhi J, Singh P, Yang DC (2017). Aluminium resistant plant growth promoting bacteria induce overexpression of aluminium stress related genes in Arabidopsis thaliana and increase the ginseng tolerance against aluminium stress. Microbiological Research 200:45-52.

Glick BR (1995). The enhancement of plant growth by free-living bacteria. Canadian Journal of Microbiology 41(2):109-117.

Glick BR, Patten CL, Holguin G, Penros DM (1999). Biochemical and genetic mechanisms used by plant growth promoting bacteria. Imperial College Press, London, United Kingdom pp 267.

Şeker MG, Şah I, Kırdök E, Ekinci H, Çiftçi YÖ, Akkaya Ö (2017). A hidden plant growth promoting bacterium isolated from in vitro cultures of fraser photinia (Photinia × fraseri). International Journal of Agriculture and Biology 19(6):1511-1519.

Hallmann J, Berg G (2006). Spectrum and population dynamics of bacterial root endophytes. In: Schulz BJ (Ed). Microbial Root Endophytes. Springer-Verlag, Berlin pp 15-31.

Hamill JD, Rounsley S, Spencer A, Todd G, Rhodes MJ (1991). The use of the polymerase chain reaction in plant transformation studies. Plant Cell Reports 10(5):221-224.

Hardoim PR, van Overbeek LS, van Elsas JD (2008). Properties of bacterial endophytes and their proposed role in plant growth. Trends in Microbiology 16(10):463-471.

Ishiga Y, Ishiga T, Uppalapati SR, Mysore S (2011). Arabidopsis seedling flood-inoculation technique: a rapid and reliable assay for studying plant-bacterial interactions. Plant Methods 7(1):32.

Jang JC, Sheen J (1994). Sugar sensing in higher plants. The Plant Cell 6(11):1665-1679.

Jang JC, Leon P, Zhou L, Sheen J (1997) Hexokinase as a sugar sensor in higher plants. The Plant Cell 9(1):5-19.

Karssen CM (1982). Seasonal patterns of dormancy in weed seeds. In: Khan AA (Ed). The Physiology and Biochemistry of Seed Development, Dormancy and Germination. Elsevier Biomedical Press, Amsterdam Netherlands pp 243-270.

Kloepper JW, Hume DJ, Scher FM, Singleton C, Tipping B, Laliberté M, … Lee L (1988). Plant growth-promoting rhizobacteria on canola (rapeseed). Plant Disease 72(1):42-45.

Kloepper JW, Lifshitz R, Zablotowicz RM (1989). Free living bacterial inocula for enhancing crop productivity. Trends in Biotechnology 7(2):39-44.

Lichtenthaler HK, Buschmann C (2001). Chlorophylls and carotenoids: Measurement and characterization by UV-VIS spectroscopy. Current Protocols Food Analytical Chemistry 1(1):F4-3.

Liu L, Kloepper JW, Tuzun S (1995). Induction of systemic resistance in cucumber against bacterial angular leaf spot by plant growth-promoting rhizobacteria. Phytopathology 85(8):843-847.

Mendes R, Pizzirani-Kleiner AA, Araujo WL, Raaijmakers JM (2007). Diversity of cultivated endophytic bacteria from sugarcane: genetic and biochemical characterization of Burkholderia cepacia complex isolates. Applied and Environmental Microbiology 73(22):7259-7267.

Moore B, Zhou L, Rolland F, Hall Q, Cheng WH, Liu YX, … Sheen J (2003). Role of the Arabidopsis glucose sensor HXK1 in nutrient, light, and hormonal signaling. Science 300(5617):332-336.

Murashige T, Skoog F (1962). A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiologia Plantarum 15(3):473-497.

Nehra V, Saharan BS, Choudhary M (2016). Evaluation of Brevibacillus brevis as a potential plant growth promoting rhizobacteria for cotton (Gossypium hirsutum) crop. Springer Plus 5(1):948.

Poupin MJ, Timmermann T, Vega A, Zuniga A, Gonzalez B (2013). Effects of the plant growth-promoting bacterium Burkholderia phytofirmans PsJN throughout the life cycle of Arabidopsis thaliana. PLoS One 8(7):e69435.

Pritchard HW, Wood JA, Mjanger KR (1993). Influence of temperature on seed germination and the nutritional requirements for embryo growth in Arum maculatum L. The New Phytologist 123(4):801-809.

Rolland F, Baena-Gonzalez E, Sheen J (2006). Sugar sensing and signaling in plants: conserved and novel mechanisms. Annual Review of Plant Biology 57:675-709.

Rook F, Corke F, Card R, Munz G, Smith C, Bevan MW (2001). Impaired sucrose-induction mutants reveal the modulation of sugar-induced starch biosynthetic gene expression by abscisic acid signaling. The Plant Journal 26(4):421-433.

Rosenblueth M, Martinez-Romero E (2006). Bacterial endophytes and their interactions with hosts. Molecular Plant-Microbe Interactions 19(8):827-837.

Ryan RP, Germaine K, Franks A, Ryan DJ, Dowling DN (2008). Bacterial endophytes: Recent developments and applications. FEMS Microbiology Letters 278(1):1-9.

Şah I (2017). Biochemical and molecular characterization of plant growth promoting putative endophytic bacterium isolated from in vitro cultures of Photinia x fraseri dress. PhD thesis, Gebze Technical University.

Sheen J (1994). Feedback control of gene expression. Photosynthesis Research 39(3):427-438.

Sukweenadhi J, Kim YJ, Choi ES, Koh SC, Lee SW, Kim YJ, Yand DC (2015). Paenibacillus yonginensis DCY84T induces changes in Arabidopsis thaliana gene expression against aluminum, drought, and salt stress. Microbiological Research 172:7-15.

Valdenegro M, Barea JM, Azcon R (2001). Influence of arbuscular-mycorrhizal fungi, Rhizobium meliloti strains and PGPR inoculation on the growth of Medicago arborea used as model legume for re-vegetation and biological reactivation in a semi-arid Mediterranean area. Plant Growth Regulation 34(2):233-240.

Weller DM, Cook RJ (1986). Increased growth of wheat by seed treatments with fluorescent pseudomonads, and implications of Pythium control. Canadian Journal of Plant Pathology 8(3):328-334.

Zhang H, Xie X, Kim MS, Kornyeyev DA, Holaday S, Paré PW (2008). Soil bacteria augment Arabidopsis photosynthesis by decreasing glucose sensing and abscisic acid levels in planta. The Plant Journal 56(2):264-273.




How to Cite

BATOOL, M., AKKAYA, Özlem, GÜL ŞEKER, M., & ÖZDEN ÇİFTÇİ, Y. (2019). Biohardening of Arabidopsis thaliana Seeds and Seedlings with Fraser Photinia Associated Bacterium (PGB_invit) in In vitro Conditions. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 47(3), 954–961.



Research Articles
DOI: 10.15835/nbha47311488

Most read articles by the same author(s)