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Vermipriming - A Noble Technology for Seed Invigouration in Rice (Oryza sativa

L.)

Article  in  The Journal of Innovation in Health Informatics · September 2016

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 SKUAST Journal of Research 18(2): 124-129; (2016)

Vermipriming - A Noble Technology for Seed Invigouration in Rice

(Oryza sativa L.)

Farooq Ahmad Khan*, Sumati Narayan, Sajad Ahmad Bhat and Rifat Maqbool

Division of Post Harvest Technology; *Division of Vegetable Science

Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar,
Srinagar-190 025, Jammu & Kashmir (India)
*e-mail: drkhan_387@skuastkashmir.ac.in

(Received 05 September 2016; accepted 29 September 2016)

ABSTRACT

Seed priming is an accepted technique to improve the seed vigour with quick
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and synchronized germination. However, success of the technique largely

depends on the methods and priming agents used. Being a rich source of plant
nutrients and other growth factors, the effectiveness of vermiwash as priming
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agent, along with other common priming substances was evaluated. Sterilized
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seeds of Shalimar rice-I were primed with PEG-6000 (0.5, 1.0 and 1.5MPa),
ZnSO4 (0.1, 0.2 and 0.3%), KH2PO4 (1.0, 1.5 and 2.0%), vermiwash (4.0, 6.0
and 8.0X dilution) and H2O where as dry seeds were taken as control. Fifty
seeds of each treatment with five replicates were sown in 14.0 cm petri dishes
lined with moist blotting paper and kept in an incubator at 25±2°C for
germination. Various priming treatments significantly improved seed
germinability and most striking figures of FGP (97.4), T50 (2.58 day), GE (72.0
%) and GV (47.4) were recorded with vermipriming followed by PEG-6000,
KH2PO4, ZnSO4 and H2O against the poorest value of FGP (81.7), T50 (4.10
day), GE (29.6 %) and GV (25.9) in nonprimed seeds. Vermipriming also
indexed maximum root and shoot dry weight (14.98 and 16.16 mg/seedling)
and length (20.2 and 26.6 mm) as well as SVI and SVII (3.03 and 455.4)
followed by KH2PO4, PEG-6000, ZnSO4, and H2O against the minimum
values of root and shoot dry weight (5.87 and 6.35 mg/seedling), length (14.3
and 18.7 mm) as well as SVI and SVII (1.00 and 269.6) noted with control.
Consequently, vermiwash should be used as priming agent for enhanced
germinability and seedling stand in rice.

Key words: Germination rate, rice, seed priming, seedling vigour, vermiwash

Rice is the most important staple food crop of the world feeding more than half of the world’s
population. Over 90% of the global rice production and consumption is centered in Asia, wherein
the world's largest rice producers, China and India, are also the world's largest rice consumers
(Anonymous, 2016). In India, the compounded annual growth rate (CAGR) of rice production has
declined from 3.62 per cent during 1981-90 to 1.90 per cent during 2000-2007 (Kannan and
Sundaram, 2011). Poor seed germination and seedling establishment is one of the major constraints
for realizing potential yield in rice. Seed priming is a recognized technique to improve the seed
vigour with quick and synchronized germination over a range of environmental conditions (Nawaz
et al., 2013; Singh et al., 2015). Seed priming is a pre-sowing technique in which seeds are
partially hydrated to a point where germination-related metabolic processes begin but radicle
emergence does not occur. The conventional way of priming seeds has been to immerse them in an
aerated solution of an osmotic material, usually polyethylene glycol (PEG). Nowadays several
 Vermipriming for seed invigouration in rice 125

variants of the usual PEG-priming technique like hydropriming (soaking seed in water),
halopriming (soaking seed in salt solutions), nutripriming (soaking seed in nutrient solutions),
matrix priming (incubation of seeds in a solid, insoluble matrix), etc. have been developed to
improve the speed and synchrony of seed germination. Vermipriming is an innovative addition to
this which involves the soaking of seeds in diluted solutions of vermiwash for a certain periods of
time and then drying the seeds up to its original weight. Vermiwash is an aqueous extract of a
column of freshly formed vermicompost and surface washings of earthworms, which contain
beneficial microorganisms, and water soluble fractions of nutrient substances and considered as a
plant tonic. As such priming of seeds with this important product is expected to provide better
results as compared to other priming techniques. Keeping these facts in mind the present
experiment was planned to evaluate the effectiveness of vermipriming along with certain
pre-existing priming techniques and appraise the usefulness of vermiwash as seed priming agent.

MATERIALS AND METHODS

The present study was carried out under laboratory conditions at the Division of Post Harvest
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Technology, SKUAST-Kashmir, Shalimar (Srinagar) during the year 2014 to establish usefulness
of vermiwash as seed priming agent for improved germination and seedling properties rice (Oryza
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sativa L.). The seed material ‘Shalimar rice -1’ having initial moisture content of 12 per cent was
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obtained from the division of Plant Breeding and Genetics, SKUAST-Kashmir. There were total 14
priming treatments which included PEG-6000 (0.5, 1.0 and 1.5MPa), ZnSO4 (0.1, 0.2 and 0.3%),
KH2PO4 (1.0, 1.5 and 2.0%), vermiwash (4.0, 6.0 and 8.0X dilution) and distilled water
(hydropriming). Dry seeds without priming were taken as control. Five grams of healthy and
uniform seeds were immersed in 10 ml of each priming solutions (25±2°C for 24 hours) and dried
back at room temperature to original moisture content (seed priming). The seeds were surfaced
sterilized in a 1:10 (v/v) dilution of commercial hypochlorite bleach for 10 min and rinsed several
times with distilled water. Fifty seeds of each treatment were sown in 14.0 cm petri dishes lined
with 5 layers of moist blotting paper. The experiment was set in an incubator (25±2°C) following
completely randomized design (CRD) with five replications. Seed germination was recorded on
daily basis according to AOSA (1990) until a constant count and final germination per cent (FGP)
was calculated. Seeds were considered to have germinated after radicle emergence. Energy of
germination (GE) was calculated as percentage of seeds germinated at 72 h (Bam et al., 2006). Time
taken to 50 per cent seed germination was calculated according to Farooq et al. (2005). Shoot and
root length of 10 randomly selected seedlings were measured after 7 days of sowing and weights
were determined by drying the seedlings at 80 ±2°C for 48 hours. The rate of germination was
estimated using a modified Timson’s index of germination velocity (GV = Σ G/t), where G is the
percentage of seed germination at one-day intervals and t is the total germination period (Tian et al.,
2014). Seedling vigour I & II (SVI and SVII) were calculated by multiplying the dry weight (g) and
length (cm) of seedlings with germination percent, respectively (Abdul-Baki and Anderson, 1973).
The data obtained were averaged and statistical analyses (ANOVA) was performed to find out the
significance of variation among the treatments while the significant difference between mean
treatments was separated using least significance difference at 5% level of probability.

RESULTS AND DISCUSSION

Various seed priming treatments grounded considerable distinctions in final germination percent
(FGP) as well as germination speeds (in terms of T50, GE and GV) of Shalimar rice-1 (Table 1).
 126 Farooq Ahmad Khan et al.

Table 1: Effect of various seed priming treatments on final germination percent and speed of
germination in Shalimar rice -1 seeds
Treatments FGP (%) T50 (Day) GE (%) GV
Nonprimed (Control) 81.7 (9.04) 4.10 29.6 (5.44) 25.9
Hydropriming (H2O) 84.9 (9.21) 3.50 34.5 (5.87) 27.5
PEG-6000 (0.5MPa) 93.8 (9.69) 3.05 49.4 (7.03) 35.1
PEG-6000 (1.0MPa) 94.0 (9.70) 2.91 58.2 (7.63) 33.2
PEG-6000 (1.5MPa) 96.7 (9.83) 3.00 50.0 (7.07) 35.4
Mean (PEG-6000) 94.8 (9.74) 2.99 52.5 (7.25) 34.6
ZnSO4 (0.1%) 87.4 (9.35) 3.10 45.2 (6.72) 30.5
ZnSO4 (0.2%) 85.3 (9.24) 3.55 35.8 (5.98) 28.9
ZnSO4 (0.3%) 84.8 (9.21) 3.40 39.0 (6.25) 25.4
Mean (ZnSO4) 85.8 (9.26) 3.35 26.7 (5.16) 28.3
KH2PO4 (1.0%) 83.7 (9.15) 3.25 41.5 (6.44) 36.8
KH2PO4 (1.5%) 86.8 (9.32) 3.15 44.0 (6.63) 39.4
KH2PO4 (2.0%) 90.1 (9.50) 3.00 50.0 (7.07) 40.5
Mean (KH2PO4) 86.9 (9.32) 3.13 45.2 (6.72) 38.9
Vermiwash (8 X) 97.0 (9.85) 2.80 66.7 (8.17) 46.5
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Vermiwash (6 X) 99.6 (9.98) 2.50 71.2 (8.44) 47.6

Vermiwash (4 X) 95.5 (9.77) 2.45 78.0 (8.83) 48.2
Mean (Vermiwash) 97.4 (9.87) 2.58 72.0 (8.48) 47.4
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C.D. (p=0.05) 0.19 0.27 0.61 5.98

Figures in parentheses are square root transformed values of the data; FGP is final germination percent; T50 is time taken
to 50 per cent germination; GE is energy of germination; GV is index of rate of germination or germination value.

Seed priming with vermiwash registered highest germination percent (97.4) followed by PEG-6000
(94.8), KH2PO4 (86.9) and ZnSO4 (85.8) against the minimum germination percent in unprimed
(81.7) as well as hydro-primed (84.9) seeds. However, FGP under vermipriming was at par with
PEG priming and in the same way difference between dry (nonprimed) and hydroprimed seeds was
found non-significant statistically. FGP of Shalimar rice-1 seeds also differed significantly among
different levels of priming treatments (except ZnSO4) wherein PEG-600 (1.5MPa), KH2PO4 (2.0%)
and vermiwash (6X) resulted in highest germination of 96.7, 90.1 and 99.6 per cent, respectively
among their respective groups. Time taken to 50 per cent germination (T 50), energy of germination
(GE) and Timson's index of germination velocity (GV) also showed the superiority of vermiwash as
evidenced by lowest T50 (2.58 day) and highest GE (72.0) as well as GV (47.4) values compared
with other priming treatments as well as control. PEG-6000 proved the second best priming
treatment in terms of T50 (2.99 day) and GE (52.5) whereas KH2PO4 was the second best in terms of
GV (38.9) values followed by KH2PO4/PEG-6000, ZnSO4 and hydro-priming, respectively in
opposed to the poorest T50 (4.10 day), GE (29.6%) and GV (25.9) values in unprimed seeds. 6X
vermiwash, 2.0% KH2PO4 and 0.1% ZnSO4 were proved as the best levels among their respective
groups by taking minimum days to 50-per cent germination (2.50; 3.00; 3.10) and showing better
energy of germination (71.2; 50.0; 45.2) as well as Timson’s index value (47.6; 40.5; 30.5).
However, minimum value of T50 (2.91 day) coupled with maximum value of GE (58.2%) was
evident with PEG-6000 (1.0MPa). Significant improvement in per cent seed germination due to
pre-soaking seed treatment with PEG (Golshani et al., 2010), Zinc (Prom-u-thai et al., 2012; Silva
and Silva, 2016), KH2PO4 (Yari et al., 2010) and vermiwash (Fathima and Sekar, 2014) has also
been reported. The improvement in seed germination attributes due to various priming treatments
may be attributed to buildup of germination-promoting metabolites (Li et al., 2010; Kamithi et al.,
2016) as well as change in water relation properties (osmotic adjustment) of the seed (Bradford,
1986; Aloui et al., 2014: Ozdemir and Sade, 2015; Hussain et al., 2016). However, an animated
improvement in FGP as well as speed of germination due to vermipriming may be attributed to the
presence of water-soluble macro- and micronutrients, growth promoting hormones, beneficial
 Vermipriming for seed invigouration in rice 127

micro-organisms, etc. which might have also caused a buildup of germination-promoting

metabolites, metabolic repair during imbibitions and osmotic regulations within the seed.
Employing of various seed priming treatments resulted in noteworthy shifts in different
growth and vigour index attributes of Shalimar rice-1 (Table 2). Seed priming with vermiwash
proved to be most effective in improving the root and shoot dry weight as well as length which
produced a seedling with 14.98 and 16.16 mg of root and shoot dry weight along with 20.2 and
26.6 mm of root and shoot length, respectively. However, unlike the germination per cent as well
as speed of germination, KH2PO4 ranked as the second best priming treatment instead of
PEG-6000 by producing root and shoot dry weight of 12.15 and 14.74 mg/plant in combination
with the root and shoot length of 19.8 and 24.00 mm, respectively as compared to 9.02 and 10.72
mg/plant of root and shoot dry weight coupled with 14.6 and 22.7 mm/plant of root and shoot
length in PEG-6000. Unprimed seeds recorded the lowest root and shoot dry weight (5.87 and 6.35
mg/plant) together with the lowest root and shoot length (14.3 and 18.7 mm/plant) in that order
followed by hydro-as well as ZnSO4-priming. Variations in the dry weight and length of the
seedling have also been observed among different levels of the same priming agents and 6X
vermiwash, 2% KH2PO4, 1.5MPa PEG-6000 as well as 0.1% ZnSO4 recorded the highest values of
seedling weight (16.72 and 17.60 mg/plant) and length (22.0 and 27.2 mm/plant) as compared to
other levels of the analogous priming agents. Our results conform to the findings of Golshani et al.
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(2010), Zarei and Sinaki (2012), Prom-u-thai et al. (2012) and Goswami et al. (2013) where in they
have reported an enhanced growth of seedlings in rice genotypes due to priming of seeds with
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PEG, KH2PO4 or zinc.

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Table 2: Effect of various seed priming treatments on growth and seedling vigour indices of
Shalimar rice -1 seeds
Treatments Root dry Shoot dry Root length Shoot SVI SVII
weight weight (mm/plant) length
(mg/plant) (mg/plant) (mm/plant)
Nonprimed (Control) 5.87 6.35 14.3 18.7 1.00 269.6
Hydropriming (H2O) 9.18 11.25 14.5 19.4 1.73 287.8
PEG-6000 (0.5MPa) 8.10 10.14 14.0 21.5 1.71 333.0
PEG-6000 (1.0MPa) 8.46 9.95 13.9 23.1 1.73 347.8
PEG-6000 (1.5MPa) 10.49 12.06 16.0 23.4 2.18 381.0
Mean (PEG-6000) 9.02 10.72 14.6 22.7 1.87 353.9
ZnSO4 (0.1%) 6.97 9.95 14.2 22.9 1.47 324.3
ZnSO4 (0.2%) 6.26 8.35 12.4 20.6 1.25 281.5
ZnSO4 (0.3%) 6.55 8.40 12.0 19.2 1.27 264.6
Mean (ZnSO4) 6.59 8.90 12.9 20.9 1.33 290.1
KH2PO4 (1.0%) 10.78 13.15 17.6 22.7 2.00 337.3
KH2PO4 (1.5%) 11.90 14.88 20.0 24.2 2.32 383.7
KH2PO4 (2.0%) 13.77 16.20 21.7 25.1 2.70 421.7
Mean (KH2PO4) 12.15 14.74 19.8 24.0 2.34 380.9
Vermiwash (8 × X) 11.75 13.35 16.6 25.5 2.43 408.4
Vermiwash (6 × X) 16.47 17.52 21.9 27.1 3.39 488.0
Vermiwash (4 × X) 16.72 17.60 22.0 27.2 3.28 469.9
Mean (Vermiwash) 14.98 16.16 20.2 26.6 3.03 455.4
C.D. (p=0.05) 1.37 1.11 3.10 3.65 0.17 13.77
SVI – seedling vigour index I; SVII – seedling vigour index II

Improved germination percentage and speed might have founded the base for further seedling
growth which led to enhanced dry weight and length of seedlings. In case of seedling vigour
indices (SVI &SVII), vermiwash again registered the appealing figures with seedling vigour
indices of 3.03 and 455.4, respectively. The higher values recorded for vermiwash were followed
 128 Farooq Ahmad Khan et al.

by KH2PO4 (2.34 and 380.9) and PEG-6000 (1.87 and 353.93) against the lowest values of SVI
(1.0) and SVII (269.6) under control aligned with ZnSO4 (1.33 and 290.1) hydro-priming (1.73 and
287.8). Variations in the seedling vigour indices among different levels of the same priming agent
followed the pattern of seedling dry weight and length. The variations in SVI and SVII due to
various priming treatments may be attributed to the FGP and seedling growth in terms of dry
weight and length as these attributes are the products of FGP and seedling dry weight (SVI) or
seedling length SVII). Corroborating findings have been reported in earlier studies by Yari
et al. (2010), Hussain et al. (2015) and Elkheir et al. (2016).

Conclusion: From the present study it may be concluded that seed priming of rice facilitates
significant improvements in seed germination and early seedling growth. Among various priming
treatments, vermipriming has proved as an excellent method of seed invigouration and should be
advocated for farmers’ adoption particularly in direct seeded crops of rice.

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