Mysore J. Agric. Sci.

, 49 (2) : 227-230, 2015

Influence of Microbial Seed Priming on Hormones Associated with Plant Growth

Promotion of French bean (Phaseolus vulgaris L.)
B. KANCHANASHRI AND M. K. SHIVAPRAKASH
Department of Agricultural Microbiology, College of Agriculture, UAS, GKVK, Bengaluru- 560 065

ABSTRACT

The present study was taken up to evaluate the effect of microbial seed priming on plant growth parameters
of French bean. The results revealed that microbial seed primed treatments exhibited increased concentration of
growth hormones like Indole Acetic Acid (IAA), Gibberellic Acid (GA) and cytokinins which in turn increased
plant growth parameters like germination percentage, seedling growth and vigour of French bean compared to
control. The treatment T12 (seeds primed with Pseudomonas fluorescens + Bacillus megaterium + Rhizobium sp.
+ Trichoderma harzianum) was found to be better among all the treatments studied.

FRENCH bean is a legume belonging to family Fabaceae
and is an important vegetable crop which serves as a
major source of protein in the diet and is also rich in
lipids. Germination is a major problem in some
leguminous crops. The inability of seeds to imbibe water
occurs in many species belonging to this family. The
imbibition (uptake) of water is usually an initial step in
the process of seed germination. Although most of
the seeds absorb water when they are placed under
moist conditions, a large proportion of seeds of some
species do not absorb water and hence called hard
or impermeable seeds. The structure called the seed
coat is responsible for the impermeability of legume
seeds (Quinlivan, 1971).
Biopriming or microbial seed priming is a treatment
followed in recent years to encourage more uniform
seed germination and induce profound changes in plant
characteristics and plants growth. This is a technology
of seed treatment that integrates biological and
physiological aspects of diseases control and plant
growth promotion. El-Mohamedy and Abdel-Baky
(2008) used biopriming as an alternative method for
enhancing germination and yield in pea plant.
Considering the problems involving the
germination of legumes, the present study was aimed
to investigate the effects of microbial seed priming on
seed germination features including germination
percentage (GP), seedling growth and vigour of French
bean.
Prior to biopriming, the priming protocol was
standardized by considering three parameters viz,
different levels of temperature (5oC, 15oC, 25oC),
soaking hours (24 h and 32 h) in different concentration
of Poly Ethylene Glycol (PEG 6000) (-2 bar, -4 bar
and -6 bar) at115 rpm and the experiments were taken
up under lab and greenhouse conditions. The
experiments that gave best results at 25oC and 32 h of
soaking in -4 bar concentration of PEG 6000 was
considered as a standard for biopriming studies. The
primed seeds were soaked in sterile distilled water with
tween 20 containing bacterial pellets and fungal spores
singly and in combinations by following the procedure
given by Raj et al. in 2004. Unprimed seeds were kept
as control.
Amylase activity of the bioprimed seeds were
estimated by following the procedure given by
Sadasivam and Manickam in 1996. A green house
experiment was taken up to evaluate the effect of
microbial seed priming on plant growth promotion in
French bean and observations on percentage seed
germination (15th day after sowing), shoot length, root
length, total biomass were recorded on 40th day after
sowing by following standard protocols. Vigour index
was calculated using the formula given by Abdul-Baki
and Anderson in 1973.
The concentration of IAA (Indole-3-Acetic acid),
GA (gibberellic acid) and Cytokinins in plant samples
were estimated by using the standard methods. The
extraction and purification of leaf sample was carried
228 B. KANCHANASHRI AND M. K. SHIVAPRAKASH
out by following the protocol given by Kettner and
Doerffling (1995). Spectrophotomertic estimation of
IAA and GA was done using the method given by
Gordon and Weber (1951) and Paleg (1965)
respectively. The cytokinins of the plant sample was
extracted following the protocol of Horgan (1978). The
bioassay was carried out following the procedure
provided by Udayakumar and Sastry (1973).
The activity of and amylase recorded was
highest in T12 which was almost equivalent to T11 and
lowest was recorded in control (Fig.1). Microbial
primed seeds when compared to control has triggered
the activity of specific enzymes such as amylase which
in turn could have increased the availability of soluble
sugars that promoted early germination and seedling
growth.
The results revealed that different treatments with
PGPR showed better germination rate compared to
unprimed control and hydroprimed seeds (Table I). The
maximum germination percentage was recorded in the
treatment T 11 (100%), which was on par with
Fig. 1. Effect of Microbial Seed Priming on Amylase activity in French bean
T12 (98.83 %) and the minimum germination percentage
of 68.37 per cent was recorded in control. In T12 the
highest root length (13.50 cm), shoot length (34.77 cm),
total biomass (23.95 g) and seedling vigour index
(4911.85) was recorded, which was on par with T11
and lowest was recorded in control. The results indicate
that the net effect of microbial seed priming with
combination of PGPRs on seed germination behaviour
and seedling growth was positive.
Concentration of IAA and GA was recorded
higher in treatment T12 (58.63 and 96.88 g/g of leaf
sample respectively), followed by T11 (56.46 and 92.17
g/g of leaf sample respectively) and minimum was
recorded in control (10.03 and 28.41 g/g of leaf sample
respectively. The cytokinin concentration was highest
in T11 (0.352 g/g of benzyl adenine) which was on
par with T12 (0.345 g/g of benzyl adenine) (Table II).
PGPRs are known to enhance seed germination
and subsequently increase the seedling vigour which
may be due to increased synthesis of hormones like
auxins, gibberellins and cytokinins. The overall effect
 INFLUENCE OF MICROBIAL SEED PRIMING ON HORMONES ASSOCIATED WITH PLANT GROWTH 229

TABLE I

Effect of microbial seed priming on germination and seedling growth in French bean

Total
Germination per Root length Root length Vigour
Treatments biomass
centage (%) (cm /pl) (cm /pl) index
(g/pl)

Control 68.37 5.01 16.47 14.81 1597.80

T2-Hydroprimed 69.97 5.98 20.83 16.11 1975.25
T3-PEG primed seeds 81.67 6.15 21.50 16.01 2343.92
T4-MSP with P. fluorescens (P.f) 82.67 8.17 23.77 17.31 2640.47
T5-MSP with B. megaterium (B.m) 79.27 8.23 24.90 17.42 2626.21
T6-MSP with Rhizobium sp.(Rhi) 82.67 6.06 27.00 19.11 2874.43
T7-MSP with T. harzianum (T.h) 79.20 8.60 23.57 15.63 2550.11
T8-MSP with P. f + T. h 86.23 8.80 28.77 19.25 2489.63
T9-MSP with B.m + T. h 88.30 9.67 29.23 19.11 3434.87
T10-MSP with Rhi + T. h 91.43 10.07 30.60 20.24 3940.63
T11-MSP with P. f + B. m + Rhi 100.00 12.07 33.65 22.38 4715.00
T12-MSP with P. f + B.m + Rhi + T. h 98.83 13.50 34.77 23.95 4911.85
S.Em 0.66 0.09 0.28 0.22 0.89
CD@ 5% 1.93 0.87 0.82 0.66 2.62

MSP : Microbial Seed Priming

TABLE II
Effect of microbial seed priming on the levels of hormones associated with plant growth promotion
in French bean
IAA GA Cytokinins
Treatments
(g / g)

Control 10.03 28.41 0.178

T2 Hydroprimed 11.13 32.96 0.189
T3-PEG primed seeds 11.44 33.24 0.195
T4-MSP with P. fluorescens (P.f) 31.91 71.29 0.212
T5-MSP with B. megaterium (B.m) 30.06 61.33 0.205
T6-MSP with Rhizobium sp.(Rhi) 37.58 78.32 0.210
T7-MSP with T. harzianum (T.h) 20.97 51.12 0.230
T8-MSP with P. f + T. h 35.44 80.08 0.296
T9-MSP with B.m + T. h 32.45 77.05 0.289
T10-MSP with Rhi + T. h 49.93 86.16 0.285
T11-MSP with P. f + B. m + Rhi 56.46 92.17 0.352
T12-MSP with P. f + B.m + Rhi +T. h 58.63 96.88 0.345
S.Em 0.92 0.93 0.006
CD @ 5% 2.69 2.73 0.018

MSP : Microbial Seed Priming

Indole-3-Acetic acid (IAA) and Gibberellic acid (GA) (g/g of leaf sample),
Cytokinins (g/g of benzyl adenine)
230 B. KANCHANASHRI AND M. K. SHIVAPRAKASH
of microbial seed priming showed a positive response
towards production of specific enzymes and growth
hormones which in turn has enhanced germination and
seedling development in French bean. Similar results
were reported by Entesari et al., (2013) where
biopriming with PGPRs was found to encourage
uniform seed germination and plant growth in soybean.
From the above study, it can be concluded that
T12 in which the seeds were primed with Pseudomonas
fluorescens + Bacillus megaterium + Rhizobium sp.
+ Trichoderma harzianum can enhance the
germination characteristics and seedling growth in
French bean and can be used in organic and sustainable
agriculture.
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