Induction Mutation of Plant Growth-Promoting Bacteria by Cold

Plasma Technology
1 1 1 2
Boonraksa Mangmee , Saisahwan Kamoi , Wasita Injai , Kanta Sangwijit
1
Demonstration school University of Phayao, University of Phayao, Phayao, 56000, Thailand.
2
Plasma Bioegineering Unit, School of Science, University of Phayao, Phayao, 56000, Thailand.

Introduction
Long-term and continuously using of chemical fertilizers affects the soil microorganisms nutrients and other living things, such as losing the balance of microorganism in soil, the erosion of nutrients in soil and the
outbreak of plant disease. So, in order to reduce the using of chemical fertilizers, PGPB, is another way to solve this problems. PGPB are the bacteria living in or on plant roots and enhance plant productivity through various
plant growth promoting activities. It is currently being used as an alternative biofertilizer in agriculture. However, the active use of PGPB sometimes encounters challenges in agricultural application, because of the
inconsistent efficiency and vitality of PGPB. Genetic engineering techniques have been applied to overcome the limitation of PGPB utilization. In this study, The efficiency of PGPB had enhancing by using cold plasma
technology which is the highly effective mutagenesis tool and environmentally friendly.

Abstract
Soil in the agriculture area contained a large amount of microorganism, which plays a crucial role in
the agriculture section. Plant growth promoting bacteria (PGPB) provide several advantages in agriculture by
play pivotal roles in nutrient acquisition and assimilation, improved soil texture, secreting and modulating
phytohormones. Furthermore, PGPB can also transform the nutrients into absorbable form for plants such as
changing phosphorus to dissolvable form and fixing the nitrogen for the plants, all leading to enhancement of
Methodology
plant growth. Improving properties of PGPB is a promising strategy for enhancing plant growth and
1. Screening of phosphate solubilizing bacteria
productivity. Aim of the study is screening PGPB and enhancing its efficiency by induction mutation using cold
plasma technology

Results Soil sampling Isolation Observation of clear zone

2. Identification of bacteria 3. Induction Mutation of bacteria

Table 1. Identification of bacteria isolates for phosphate solubilization
isolates Phosphate solubilization pathogenesis
AF03 + - Bacillus subtilis
AF15 + + Burkholderia ambifaria
AF28 + + Burkholderia ambifaria
UP01 + + Burkholderia contaminans
Bacterial Identification by 16s rDNA Sequencing
UP07 + - Bacillus subtilis
UP13 + - Bacillus subtilis
UP46 + + Staphylococuus hominis
Induced mutation
by Cold Plasma Technology
Table 2. pH measurement at different days of cultivation Analyzing through BLAST

Sample 3 Days 5 Days 7 Days

4. Screening of bacterial mutant 5. Investigation of plant growth
PVK broth 5.86 5.96 5.91

UP13 (control) 5.24 5.38 5.08

UP13-3M 15 5.28 5.02 4.97

UP13-3M 16 5.11 4.93 4.84

UP13-3M 18 5.26 5.17 5.05

UP13-10M 1 5.07 4.85 4.66

Screened mutant bacteria and measure pH
UP13-10M 2 5.09 5.15 4.91

UP13-10M 3 5.22 4.90 5.00

UP13-10M 4 5.27 4.99 4.94

UP13-10M 14 5.21 5.13 5.02 Acknowledgements

This project was supported by Science Classroom in University Affiliated School (SCiUS). The funding of SCiUS is
provided by Ministry of Higher Education, Science, Research and Innovation. This extended abstract is not for
Table 3. Investigation of plant growth by measure after 14 days of planting citation.
Sample Width of leaves Height (cm) Root length (cm) Number of seedling
(cm) (percentage of
germination)
Reference
H2 O 0.4960.134 5.783 1.066 0.900 0.141 18 (60%)
1. Glick BR. Plant growth-promoting bacteria mechanisms and applications. Scientifica 2012;2012.
2. Naito K, Kusaba M, Shikazono N, Takano T, Tanaka A, Tanisaka T and Nishimura M. Transmissible and nontransmissible mutations
induced by irradiating Arabidopsis thaliana pollen with gamma-rays and carbon ions. Genetics 2005; 169, 9-881.
Control 0.439 0.108 5.785 0.971 2.550 0.353 27 (90%) 3. Gopinath KP, Murugesan S, Abraham J and Muthukumar K. Bacillus sp. mutant for improved biodegradation of congo red: Random
mutagenesis approach. Bioresour. Technol. 2009; 100, 300.
4. Timmusk S, Behers L, Muthoni J, Muraya A and Aronsson AC. Perspectives and challenges of microbial application for crop improvement.
Frontiers in Plant Science 2017.
Mutant 0.414 0.126 5.971 0.860 3.900 2.546 21 (70%)
5. Ubonrat J, Moonmangmee N, Kunnajak N and Moonmangmee D Isolation and screening of plant growth promoting bacteria (PGPB) from
rice (Oryza sativa) and rhizosphere soil. Science and echnology RMUTT 2018;8:190-204.