INDUCING THE VIABILITY
OF DETERIORATED JATROPHA SEEDS THROUGH MATRICONDITIONING TECHNOLOGY AND Pseudomonas fluorescens AS BIOLOGICAL
AGENT
Sundahri,
Tofan Mursyidto, Tri C. Setiawati, Hardian A. Susilo, Ali Wafa
Faculty
of Agriculture, Universitas Jember, Indonesia
Email: [email protected], [email protected],
[email protected], [email protected],
[email protected]
|
KEYWORDS Jatropha; matriconditioning; Pseudomonas
fluorescens; seeds |
ABSTRACT Jatropha curcas is a prospective vegetable oil or biodiesel and
various derivative products as well as phytoremediation (plants cleaning the
environment) and phytomining (plants that mine metals such as gold, nickel
etc.). However, the commodity often experiences problems in the supply of
seeds. The seed used usually has decreased their viability after going
through a certain storage period. Therefore, matriconditioning technology and
the application of the biological agent Pseudomonas fluorescens were
used to increase the viability and vigor of these seeds. The aim of the study
was to determine the interaction between the duration of the matrioditioning
treatment and the application of Pseudomonas fluorescens to increase
the viability and vigor of Jatropha seeds and to determine the optimal
duration of the matrioditioning treatment and the dosage of Pseudomonas
fluorescens in increasing the viability and growth performance of sprouts
and seedlings. Field experiments would be carried out in the greenhouse of the
Faculty of Agriculture, Universitas Jember. This experiment was arranged
using a factorial design (two factors) with a randomized block design (RBD)
repeated 3 times. The result showed that the interaction effect of matriconditioning time and dose of Pseudomonas
fluorescens was significantly different on the observed variables of
growth speed, vigor index and T50. Combination of 24 hours matriconditioning
treatment and dose of Pseudomonas fluorescens 100
ml l-1 produced the best treatment on variables representing seed
vigor (growth rate and vigor
index). |
INTRODUCTION
Jatropha curcas is a prospective plant, as a source of
biofuel and various kinds of its derivative products. For the development, the
seeds of the plant experience deterioration after going through a storage
period and decrease in quantity due to biotic factors (Rustam &
Audina, 2018). To induce the viability
of Jatropha seeds which have experienced deterioration, a special technology or
seed treatment is needed, in order to increase the vigor and viability of the
seeds so that they can be used as planting material. Invigoration technique is
an example for seed treatment before planting which aims to increase the value
of seed viability and vigor, indicated by increasing germination and seed
performance. An invigoration technique that has been proven successfully in
increasing the viability of some seeds is matriconditioning technology. The
induction is a controlled hydration treatment depending on the moist solid
media used, has a low matrix potential and negligible osmotic potential (Khan, 1992).
Invigoration
of matriconditioning can stimulate seed metabolism in germination; on the other
hand, the radicle emergence can be
delayed and is proven to improve seed performance (Sucahyono, 2013). Seeds that have been given the
matriconditioning treatment have metabolic conditions in the seeds that are
ready to germinate, so that the percentage of germination power and the speed
of growth of sprouts increases. In a research on soybean plants, this technique
was able to increase seed viability with increased germination parameters and
growth rates (Priyanto, 2017). In addition, the combination of
invigoration with the addition of biological agents to nyamplung seeds
can increase seed viability by up to 42% (Yuniarti,
2020).
The biological agents commonly used as seed
treatments are from the PGPR (Plant Growth Promoting Rhizobacteria) group, one
of which is Pseudomonas fluorescens. The secondary metabolites produced
by Pseudomonas fluorescens are useful for stimulating seed germination,
in the form of the hormones auxin indole acetic acid (IAA) (Istiqomah et
al., 2017), cytokinins (Kapoor &
Kaur, 2016) and gibberellins (Tefa, 2015). Siderophore compounds as bioprotectants
against pathogens are also produced by Pseudomonas fluorescens from
metabolic processes. The use of the biological agent Pseudomonas fluorescens
on the invigoration of testa hard (teak) type seeds was able to increase
the maximum growth potential and germination rate by 32.65% (Afa, 2008).
Based on the above background,
matriconditioning technology and the application of the biological agent Pseudomonas
fluorescens have to be studied to induce Jatropha seeds whose decreased
viability, especially in study of the length of matriconditioning treatment and
the density of Pseudomonas fluorescens in order to increase optimally
the viability of Jatropha seeds. In fact, each seed has a different character
and sensitivity to the seed treatment above. Therefore, this study aims to
determine the effect of the interaction between matriconditioning duration and Pseudomonas
fluorescens dose and to determine the optimal matriconditioning duration
and Pseudomonas fluorescens dose to increase the viability and growth
performance of Jatropha shoots and seedlings.
RESEARCH
METHOD
Place and Time
Field research was carried out in the
greenhouse of the Faculty of Agriculture, The University of Jember. Laboratory
research activities are carried out at the Laboratory of Genetics, Microbiology,
and Biotechnology, Faculty of Teaching and Education; Soil Chemistry and
Fertility Laboratory; and the Seed Laboratory of the Faculty of Agriculture,
University of Jember. The research time starts from June to November 2022.
Materials
and Tools
The important ingredients to be used are the
Jet 1 Agribun variety from the Research Institute for Sweeteners and Fiber
Plants (BALITTAS). Pseudomonas fluorescens isolate was obtained from the
Observation Laboratory of Food Plant Pest and Horticultural Pests in Tanggul,
Jember. While important tools needed in testing free fatty acids, electrical
conductivity, Pseudomonas fluorescens culture and testing bacterial
density.
Experimental design
Two-factor research had been carried out
using Factorial RAK with 3 repetitions. The first factor was the long
matriconditioning treatment consisting of 4 levels, namely 12 hours, 24 hours,
36 hours and 48 hours. The second factor was the density of Pseudomonas
fluorescens, consisting of 3 levels, namely 0 (without Pseudomonas
fluorescens application), dose of 100 and 200 ml.l-1.
Research Implementation
Seed
Deterioration Testing
Analysis of free fatty acids and electrical
conductivity (EC) was carried out to find out the deterioration of the seeds.
Analysis of free fatty acid levels used the method used by Hasanuddin (2010). The EC test was based on the method used by
Zanzibar (2016).
Culture of the biological agent Pseudomonas
fluorescens
Fluorescence test was carried out to see that
the isolate was a bacterium (Pf) Pseudomonas fluorescens. The bacterial
isolate of Pseudomonas fluorescens introduced from the Laboratory of
Monitoring Pests of Food Plant Diseases and Horticulture of the Embankment.
Then, the bacterial culture was carried out using NB media (Nutrient Broth).
The complete method was adopted from the method used by Scales et al. (2014).
Determination of Bacterial Density
The method used is TPC (Total Plate Count)
spread plate to determine the density of Pseudomonas fluorescens bacteria
(Yunita et al., 2021).
Matriconditioning
Treatment and Application of Pseudomonas fluorescens
Mixing
of materials was carried out on plastic clips for each treatment and stored for
12 hours, 24 hours, 36 hours and 48 hours at room temperature. Each treatment
units used a comparison of seeds, husk charcoal media, bacterial
solution/suspension (97.6 g : 48.8.1 g : 48.8 ml). Sterilized roasted husks and
seeds were added according to the comparison on the plastic clip labeled for
each treatments. The treatments consisted of a control
(without bacteria) and a bacterial solution prepared and mixed with 1:10
distilled water (density 100 ml.l-1)
as much as 200 ml l-1 and a bacterial solution prepared as much as
200 ml. The solution was poured according to each treatments, and stored
according to the matriconditioning old treatment level.
Planting
Planting
was carried out after the matriconditioning treatment and the application of
the biological agent Pseudomonas fluorescens were completed. The media
attached to the seeds is cleaned using clean water. Sand that is ready to use
before planting is watered so that the media becomes moist. The seeds are
planted to a depth of 2-3 cm in an upright position.
Observational Variables
Preliminary
observation variables that would be carried out to assess the deterioration of
the seeds used as above include fatty acid levels and electrical conductivity;
while the observations after the experiment were as follows:
1. Growth rate was calculated based on the ISTA
standard (ISTA, 2014):
2. The vigor index was calculated based on the Ilyas (2012) method with the formula:
3. Simultaneous growth is calculated by the formula:
4. Germination capacity (GC) (not
attacked by pests or diseases) was calculated based on the ISTA standard (2014)
with the formula:
5. Seed growing time was calculated by the formula:
WTB
6. Maximum potential growth was
calculated based on normal sprouts and abnormal sprouts until the end of the
observation, which was 14 days of observation (Sutopo, 2010).
7. Sprout height
Measurement
of the height of dicotyledonous sprouts was based on the height of the tip of the
growing point to the base of the stem.
8.
Normal sprout dry weight
Normal
dry weight of sprouts was a measure of potential viability which described the
amount of food reserves available so that when conditions were in an
appropriate environment they could grow and develop properly (Sadjad, 1989).
10. Abnormal sprouts (infected by pests or diseases)
This
parameter was used to find out whether the given treatment affected the
percentage of abnormal seeds or infected diseases that grew significantly.
The research data were statistically analyzed
using ANOVA. If the treatment had a significant effect, the data were further
analyzed using Duncan's Multiple Range Test (DMRT) with a 95% confidence level as
well as correlation and regression analysis to determine the optimal level.
RESULTS AND DISCUSSION
Research Results
Seed biochemical analysis tests were carried
out to obtain information on seed viability estimation before being used as
material for invigoration experiment. Estimation of seed viability was carried
out by finding the free fatty acid (FFA) and electrical conductivity (EC)
values of the seeds.
Results of free fatty acid test analysis on several
seed lots
Analysis
of free fatty acid levels from the extraction of seed fat could be used as an
indicator of seed viability. The results of the analysis on seeds with varying
viability yielded different percentages of seed fat content.
Figure 1. Results of analysis of the fat content of jatropha seeds
The
percentage of free fatty acids from the yield could be used as an illustration
of the viability of jatropha seeds. The longer the seeds are stored, the free
fatty acids of the seeds will increase (Table 1).
Table 1. Result of Jatropha seed
free fatty acid test
|
Seed Lots |
% Fat |
% FFA |
|
80% GC Seeds |
38,9 |
2.40 |
|
GC Seeds 50.67% |
37,7 |
4.94 |
|
GC Seeds 0% |
32,4 |
16.07 |
The content of fatty acids in seeds could be
used as a determinant of the estimation of seed viability and vigor values.
Free fatty acid titration results on seeds using KOH showed a change in the
colour of the fat to pink (Figure 2).
Figure 2. (a) 0% GC of seeds (b) 50.67% GC of seeds (c) 80% GC of seeds
The
use of KOH in the jatropha seed fat titration process can neutralize the free
fatty acids as indicated by the change in the colour of the fat to pink. The
more KOH solution needed to neutralize 1 g of fat, the lower the seed
viability. Jatropha seed viability and vigor decreased more quickly than other
seeds with carbohydrate (starch) content, due to the high content of
unsaturated fat in jatropha seeds. The fat undergoes oxidation which would
affect the viability and vigor of the seeds. The occurrence of the oxidation
process of unsaturated fats produced free radicals in the seeds which trigger a
decrease in the viability and vigor of jatropha seeds.
Oil content in
seeds was influenced by many external factors including the seed production
process, storage period, and the environmental seed storage. The influence of
internal factors that affected the fat content contained in jatropha seeds is
plant genetics. The change of fat into free fatty acids would be quickly
damaged if the seeds were not immediately used for germination, so that the
available energy reserves in the seeds were reduced. The available energy
reserves in the seeds affected the value of seed viability and vigor.
The results of
the analysis of the electrical conductivity of jatropha seeds
The EC test method is a simple test for estimating seed quality that
has been suggested by ISTA (International Seed Testing Association) for several
types of seeds. The electric conductivity test has the principle that seeds
will experience a reaction when electrified which is affected by cell membrane
leakage due to changes in seed permeability. Seeds that have low viability and
vigor will release a lot of elements present in the seeds in the form of
elements K, Cl, amino acids and sugars. These elements will accumulate in the
liquid used to soak the seeds. Seeds with lower viability and vigor during the
EC test will show higher conductivity values.
The
conductivity values of several seed lots presented in (Table 2) can be used
as an illustration of the viability of jatropha seeds by looking at the leakage
of the cell membranes in the seeds.
Table 2. Results of the electrical conductivity (EC)
test of jatropha seeds
|
Seed Lots |
Electrical
Conductivity (µmhos cm-1g-1) |
|
80% GC of seeds |
44.88 |
|
GC of seeds 50.67% |
56,38 |
|
GC of seeds 0% |
78,46 |
The difference in the conductivity value indicates
that the longer the jatropha seeds are stored the conductivity value increases.
The conductivity test results of Jatropha seeds used as research planting
material were 56.38 µmhos cm-1g-1. The seed
lot with 80% viability has a conductivity value of 44.88 µmhos cm-1g-1, lower 11.5 µmhos cm-1g-1 compared to the
GC of seed lot of 50.67%. Lower 33.58 88 µmhos
cm-1g-1compared to 0% GC of seed lots.
The higher the conductivity value of Jatropha seeds
the lower the viability of the seeds, indicating that the seed lot had cell
leakage and low permeability. Seeds that experienced deterioration showed
decreased cell membrane integrity and cell wall permeability. Cell leakage in
seeds occured as a result of decreased cell membrane integrity, resulting in
the dissolving of food reserves in seeds when soaking in imbibition solutions.
The decrease in the integrity of the cell membrane was affected by seed storage
time, followed by a decrease in the value of seed viability and its vigor. The
decrease in the integrity of the cell membrane was caused by the respiration
activity of the seeds and the effect of free
radicals. Free radicals caused damage to the cell membrane that interferes with
electron transport, spreading free electrons that join with free radicals
resulting in damage to the integrity of the germ cell membrane.
Damage to cell membranes and decreased cell integrity in seeds have
an impact on differences in seed metabolic activity during imbibition.
Uncontrolled hydration during seed imbibition caused cell membrane damage. The
imbibition process on seeds that experience deterioration with the
characteristic of cell membrane damage could be given controlled hydration, so
that germination performance could be improved.
The accuracy of the estimation of the electrical
conductivity test was affected by the moisture content of the seeds, the
temperature of immersion in the liquid, the duration of immersion, the dirt of
the seeds and the mechanical damage to the seeds. The permissible moisture
content was in the range of 11-17%, with a soaking time of 16-24 hours, comes
from pure seed lots and was not mechanically damaged. Temperature, soaking
time, mechanical damage and seed impurities could affect the conductivity value
of the tested seeds, so it was necessary to standardize the EC test method on
jatropha seeds.
Experiment Results
Based on
the results of analysis of variance withα= 5%, the F-value was obtained to
determine the effect of matriconditioning invigoration and the application of
the biological agent of Pseudomonas fluorescens on various observed
variables on jatropha seeds in Table 3.
Table 3. Summary of F-count results of the analysis
of variance of the observation variables
|
No |
Observational
Variables |
Compute F Value |
|||
|
|
Old Matriconditioning (A) |
Pseudomonas fluorescens (B) |
Interaction (A×B) |
||
|
1. |
Growth Rate |
14,185 ** |
62,030 ** |
2,712 * |
|
|
2. |
Vigor Index |
14,698 ** |
61,899 ** |
2,726 * |
|
|
3. |
Simultaneous
Growth |
3,867 * |
12,419 ** |
1.133 ns |
|
|
4. |
Germination
Capacity |
3,867 * |
12,419 ** |
1.133 ns |
|
|
5. |
T50 |
429,302 ** |
171,963 ** |
65,705 ** |
|
|
6. |
Maximum Growth
Potential |
2,825 ns |
7,596 ** |
0.908 ns |
|
|
7. |
Plant Height |
0.078 ns |
0.429 ns |
0.073 ns |
|
|
8. |
Normal Sprout
Dry Weight |
0.443 ns |
0.067 ns |
0.298 ns |
|
|
9. |
Abnormal Sprouts |
1.698 ns |
1.070 ns |
0.920 ns |
|
(*) =
Significantly different
(**) =
Very significantly different
(ns) =
Insignificantly different
Based on Table 3, it showed that there was an
interaction in the old treatment matriconditioning invigoration and the
biological agent of Pseudomonas fluorescens on the observed variables of
growth rate, vigor index and T50. The effect of a single factor in the
matriconditioning treatment showed significantly different values for the
variables of simultaneity of growth and germination. The matriconditioning
treatment had no significant effect on the maximum growth potential, plant
height, dry weight of normal and abnormal sprouts. The effect of a single
factor treatment of Pseudomonas fluorescens was highly significant to
the variable of growth simultaneity, germination capacity and maximum growth
potential. The treatment of Pseudomonas fluorescens had no significant
effect on plant height, dry weight of normal and abnormal sprouts.
Results of Regression
Correlation Analysis between Observational Variables
Correlation
is a measure of the level of closeness and direction of the relationship between
two variables (R). The regression coefficient is a measure of the magnitude of
change in one variable related to one experimental unit and another variable.
Correlation analysis was carried out on the growth rate, vigor index and T50
variables on the jatropha seed germination variable. Correlation and regression
analysis between variables could be seen in Table 4.
Table
4. Summary of R values and t-counts of regression coefficients and
correlations
|
No. |
Observational variable |
r |
t-count |
|
|
X |
Y |
|||
|
1 |
Growth Speed |
Germination |
0.95 |
9.43 * |
|
2 |
Vigor Index |
Germination |
0.95 |
9.40 * |
|
3 |
T50 |
Germination |
0.43 |
1.48 ns |
The
results of regression analysis and correlation based on the T test (comparison
of t-count with t-table shows that there was a significant correlation between
the growth rate and vigor index variables on seed germination, t-count>
t-table. There was no significant correlation between the variable of T50 on
jatropha seed germination (t-count < t-table), the value of t-table compared
to the t-count used was 2.22814. The interpretation of the correlation
coefficient (r) as follows: 0.00-0.20 very low, value 0.20-0.40 low, value
0.40-0.70 sufficient, high correlation 0.700-0.90, value> 0.90 very high.
Relationship between growth speed
and seed germination
The
relationship between the observed variable growth rate and seed germination
from the results of correlation and regression analysis was presented in Figure
3.
Figure 3.
Correlation of growth rate variable to seed germination capacity
Figure 3 showed that the growth rate and germination
variables had a unidirectional relationship with a very high degree of
closeness. The regression equation shown was y = 5.5931x + 19.307 which meant
that every increase in growth rate of 1%/etmal would be followed by an increase
in seed germination capacity of 5.5931%. This correlation indicated that any
increase in seed vigor, represented by growth rate, would be followed by an
increase in viability, represented by the variable seed germination. Seed
viability and vigor were a unit of seed quality which had the same cycle graph
forming a sigmoid-shaped curve. The curve meant that the decrease in quality
that occurs in seeds was preceded by a decrease in vigor and was followed by
seed viability.
Correlation
between vigor index variables and seed germination
The
relationship between the observation variable vigor index and seed germination
from the results of the correlation and regression analysis was presented in
Figure 4.
Figure 4. Correlation of the vigor
index variable on seed germination
Figure
4 showed that the vigor index variable on seed germination had a unidirectional
relationship with a very high degree of closeness. The regression equation
shown was y = 0.7942x + 19.452 which meaned that every 10% increase in the
vigor index would be followed by a 7.94% increase in seed germination. This
correlation indicated that any increase in seed vigor represented by the vigor
index would be followed by an increase in viability represented by the variable
seed germination. The increase in seed viability in the form of germination had
a close relationship with the value of the vigor index, so that the high value
of germination could be known from the first count germination performance.
Effect
of Interaction of Matriconditioning and Pseudomonas fluorescens in
Increasing Jatropha Seed Viability and Vigor
Based on analysis of variance of all observed variables, it was shown
that the effect of the interaction between the duration of matriconditioning
and the dose of Pseudomonas fluorescens was significantly different from
the observed variables of growth velocity, vigor index and T50.
Growth Rate
(Kct)
The effect of matriconditioning invigoration time and biological agent Pseudomonas
fluorescens on growth speed variables was presented in Table 5.
|
Pf dose (mll-1) |
Matriconditioning (hours) |
|||
|
12 |
24 |
36 |
48 |
|
|
0 |
5.14 ± 0.57 |
5.7 1 ± 0.00 |
6.82 ± 0.57 |
4.95 ± 0.66 |
|
ab |
ab |
c |
a |
|
|
100 |
7.24 ± 0.33 |
8.76 ± 0.66 |
9.09 ± 0.98 |
6.29 ± 0.57 |
|
c |
d |
d |
bc |
|
|
200 |
5.14 ± 0.57 |
5.33 ± 0.33 |
5.49 ± 0.66 |
5.71 ± 0.66 |
|
ab |
ab |
ab |
a |
|
The numbers followed by
the same letters were not significantly different according to Duncan's test at
the 5% level of significance.
The relationship between the duration of matriconditioning treatment
and the dose of Pseudomonas fluorescens on the growth rate (Kct) of
jatropha seeds was also presented in Figure 5.
The combination treatment that produced the highest Kct occurred in the
36-hour matriconditioning treatment at a dose of Pseudomonas fluorescens 100 ml l-1 for9.09%/etmal, but not
significantly different from the 24-hour matriconditioning combination treatment
at the dose of Pseudomonas fluorescens 100 ml
l-1 of 8.76%/etmal. The same
matriconditioning time at the level of 24 hours produces mark The Kct was significantly different at the
doses of Pseudomonas fluorescens. Same dose of Pseudomonas fluorescens 100 ml l-1 yield value Kct which was not
significantly different at 36 hours and 24 hours matriconditioning time.
The combination of treatments that produced the lowest growth rate
values occurred in the 48-hour long matriconditioning treatment, at a dose of
0 ml of Pseudomonas fluorescens and 200 ml l-1.The
best treatment suggested in this experiment was based on the parameter of
increasing Kct, namely the combination of 24 hours of matriconditioning and
doses of Pseudomonas fluorescens 100 ml l-1.The value of growth speed was obtained from observing the
growth of Jatropha seed sprouts in units of the percentage of normal sprouts
per etmal. The higher the value of Kct produced in this study meant that
the seed vigor had increased.
Vigor Index (VI)
The effect of matriconditioning invigoration time and biological agent Pseudomonas
fluorescens on the vigor index variable was presented in Table 6.
|
Pf dose (ml.l-1) |
Matriconditioning (hours) |
|||
|
12 |
24 |
36 |
48 |
|
|
0 |
36±4.00 |
40±0.00 |
48±4.00 |
34.67±4.62 |
|
ab |
ab |
c |
a |
|
|
100 |
50.67±2.31 |
61.33±4.62 |
64±6.93 |
44±4.00 |
|
c |
d |
d |
bc |
|
|
200 |
36±4.00 |
37.33±2.31 |
38.67±4.62 |
34.67±4.62 |
|
ab |
ab |
ab |
a |
|
The numbers followed by
the same letters were not significantly different according to Duncan's test at
the 5% level of significance.
Display of the relationship between matriconditioning duration and Pseudomonas
fluorescens dose on the vigor index value of Jatropha seeds was presented
in graphical form which could be seen in Figure 6.
The combination treatment that produced the highest vigor index was
found in the 36-hour matriconditioning treatment at a dose of Pseudomonas
fluorescens 100 ml l-1 for 64%
but not significantly different from the 24-hour matriconditioning combination
treatment at the dose of Pseudomonas fluorescens 100 ml l-1 for 61.33%. The same matriconditioning time
at the 24 hours level resulted in significantly different vigor index values
at the doses of Pseudomonas fluorescens. Same dose of Pseudomonas
fluorescens 100 ml l-1 yield value vigor
index which was not significantly different at 36 hours and 24 hours
matriconditioning time. The combination of treatments that produced the lowest
vigor index value occurred in the 48-hour long matriconditioning treatment, at
a dose of 0 ml of Pseudomonas fluorescens and
200 ml l-1. The best treatment suggested in this experiment
was a combination of 24 hours of matriconditioning and doses of Pseudomonas
fluorescens 100 ml l-1.
T50 (50%
Seed Growing Time)
The effect of matriconditioning invigoration time and the biological
agent Pseudomonas fluorescens on variable T50 was presented in Table 7.
|
Pf dose (ml.l-1) |
Matriconditioning (hours) |
|||
|
12 |
24 |
36 |
48 |
|
|
0 |
11.92±0.00 |
9.68±0.00 |
7.04±0.00 |
0.00±0.00 |
|
d |
c |
ab |
e |
|
|
100 |
7.95±1.02 |
7.53±0.89 |
6.31±1.48 |
0.00±0.00 |
|
b |
b |
a |
e |
|
|
200 |
10.53±0.00 |
0.00±0.00 |
0.00±0.00 |
0.00±0.00 |
|
c |
e |
e |
e |
|
The numbers followed by
the same letters were not significantly different according to Duncan's test at
the 5% level of significance.
Display of the relationship between matriconditioning duration and Pseudomonas
fluorescens dose on the vigor index value of Jatropha seeds is presented in
graphical form which could be seen in Figure 7.
The combination treatment that produced the fastest T50 occurred in the
36-hour matriconditioning treatment at a dose of Pseudomonas fluorescens 100 ml l-1. The T50 value of the
interaction effect on the 36-hours matriconditioning treatment and the combined
dose of Pseudomonas fluorescens 100 ml l-1
with a value of 6.31 was better than the old matriconditioning hour treatment
without Pseudomonas fluorescens administration, but there was no
significant difference between the two treatments. The same matriconditioning
time at the 36 hours level resulted in a T50 value that was not significantly
different at the dose of Pseudomonas fluorescens 100 ml l-1 and without Pseudomonas fluorescens. Same dose of Pseudomonas fluorescens 100 ml l-1 generate
valueT50 which was significantly different in the old matriconditioning
treatment. A value of 0 (zero) on the results indicated that the combination of
these treatments did not achieve a minimum germination of 50%. The best
treatment suggested to increase the T50 value in this experiment was a
combination of 36 hours of matriconditioning without the application of Pseudomonas
fluorescens.
The
Effect of a Single Factor of Matriconditioning Old Treatment in Increasing
Viability and Vigor of Jatropha Seeds
Simultaneous
Growth (Kst)
The results of the analysis of variance in (Table 3) showed that the
matriconditioning duration had a significant effect on the growth simultaneity
value (Kst) of Jatropha curcas seeds. Based on the DMR follow-up test,
the highest Kst value for the matriconditioning duration was achieved at the 36
hours level, namely 58.22%. Further test results showed that the
matriconditioning duration among 12 hours, 24 hours and 36 hours resulted in a
Kst that was not significantly different, respectively 55.11%, 56.44%, 58.22%.
The Kst value of the 12, 24 and 36 hours
matriconditioning duration was significantly different from the 48 hour
matriconditioning treatment which had the smallest Kst value of 47.11%. The
negative impact of the matriconditioning treatment occurred at the 48 hours
level resulting in a Kst value of 47.11%, which was followed by a decrease in
the value of seed germination.
Figure
8. The effect of long matriconditioning treatment on the uniformity of growing
Jatropha seeds
The
results of the orthogonal polynomial test produce a regression equation (y) =
-x2 + 1.111x + 44.222 with a coefficient of determination (R²) = 0.877. Based
on the regression equation, it showed that the optimal value for the 25.72
hours long matriconditioning treatment resulted in an optimal Kst of 58.57%.
The 24 hour matriconditioning time was the best time to affect the simultaneity
of Jatropha seed growth, although it was not significantly different from the
12 hour and 36 hour matriconditioning time. The matriconditioning time of 24
hours was close to the time that produces the optimal value of growing
simultaneity. The high value of growing simultaneously indicated that
matriconditioning was able to stimulate seed metabolism so that it could grow
simultaneously in the planting area.
Germination
Capacity (GC)
The prolonged matriconditioning treatment had a significant effect on
the germination rate of Jatropha seeds as shown by the results of the analysis
of variance in (Table 3). Based on the DMRT follow-up test, the highest
germination capacity of matriconditioning was achieved at the 36 hours level,
namely 58.22%. Further test results showed that the matriconditioning duration
between 12 hours, 24 hours and 36 hours resulted in a GC that was not
significantly different respectively 55.11%, 56.44%, 58.22%. The increase in
the GC value for the matriconditioning time of 12 hours, 24 hours and 36 hours
against the GC test value without treatment was 4.44%, 5.77%, 7.55%,
respectively. The GC values of the 12, 24 and 36 hours
matriconditioning duration were significantly different from the 48 hour
matriconditioning treatment which had the smallest GC value of 47.11%.
Based on the DMRT test on the observation variable germination had the
same value as the simultaneity of growth, because the addition of sprouts ends
in calculating the simultaneity of growth (11 DAP). The results obtained from
the study, although not significantly different at the level of
matriconditioning time of 12 hours, 24 hours and 36 hours, proved that matriconditioning
was able to increase the germination value of jatropha seeds. The increase in
GC values in this study is in line with Mariani and Andi (2021), research on
soybean seeds resulted in an increase in GC values of up to 13% compared to
no matriconditioning treatment. The 48-hours matriconditioning treatment caused
the GC value to decrease by 3.56% compared to the GC test value without
treatment. The increase in GC value was an indicator that prolonged
matriconditioning treatment could increase seed viability. The relationship
between matriconditioning and long-term germination of Jatropha seeds could be
seen in Figure 9.
Figure 9.
Effect of long matriconditioning treatment on jatropha seed germination capacity
The results of the orthogonal polynomial test produced a regression
equation (y) = -x2 + 1.111x + 44.222 with a coefficient of determination (R²) =
0.877. Based on the regression equation, it showed that the optimal value for
the long matriconditioning treatment was 25.72 hours, resulting in an optimal
GC of 58.57%. The 24 hours matriconditioning time was the best time to affect
the simultaneity of Jatropha seed growth, although it was not significantly
different from the 12 hour and 36 hours matriconditioning time. The
matriconditioning time of 24 hours was close to the time that produces the
optimal value of growing simultaneity.
Effect of Single Factor Treatment of Pseudomonas
fluorescens Dosage in Increasing Jatropha Seed Viability and Vigor
Simultaneous
Growth (Kst)
The results of the analysis of variance in (Table 3) showed that the
treatment dose of Pseudomonas fluorescens had a significant effect on
the growth simultaneity (Kst) value of Jatropha curcas seeds. Based on the DMRT
follow-up test, Pseudomonas fluorescens dose treatment 100 ml l-1 produced the highest Kst
value, compared to other treatments with a Kst value of 63%. Kst value of Pseudomonas
fluorescens dose treatment100 ml l-1 significantly
different from the control treatment and at the dose of Pseudomonas
fluorescens 200 ml l-1 which
had a GC value of 50.33% and 49.33% respectively. Kst value of Pseudomonas
fluorescens dose treatment100 ml l-1 significantly
different with dose 200 ml l-1 and control. Pseudomonas fluorescens dose
treatment100 ml l-1 was able to increase
the Kst value by 12.67% compared to the control.
Pseudomonas fluorescens dose treatment200
ml l-1 produced values that were not significantly
different from the control, and produced the smallest Kst value. Pseudomonas
fluorescens dose treatment 200 ml l-1 negatively
affected the Kst value, decreasing the Kst value by 1% compared to the Kst
control. The decrease in the value of the simultaneity of growth was suspected
to be the dose of Pseudomonas fluorescens 200
ml l-1, had a toxic effect on seeds in the germination process. The
relationship between the dose of Pseudomonas fluorescens and the
simultaneous growth of Jatropha seeds could be seen in Figure 10.
The results of the orthogonal polynomial test produced a regression
equation (y) = -x2 + x + 50.333 with a coefficient of determination (R²) = 1.
Based on the regression equation, it showed that the optimal value was at the
treatment dose of Pseudomonas fluorescens 99.35 ml
l-1 produces an optimal value of Kst 63.16%. Pseudomonas
fluorescens 100 dose ml l-1 be
the best dose in affecting the simultaneous growth of Jatropha seeds compared
to other doses. Dose 100 ml l-1
Pseudomonas fluorescens approached the time that produced the optimum value
of the growing synchrony of Jatropha seeds. According to Sutariati et al.
(2014), the application of Pseudomonas fluorescens used in
matriconditioning invigoration had an effect of increasing the Kst value by 19%
compared to matriconditioning without Pseudomonas fluorescens on upland
rice seeds.
Germination
Capacity (GC)
The results of the analysis of variance in (Table 3) showed that the
dose of Pseudomonas fluorescens had a significant effect on the
germination rate of jatropha seeds. Based on DMRT follow-up test with Pseudomonas
fluorescens dose treatment 100 ml l-1 produces
the highest DB value, compared to other treatments with a germination rate
value of 63%. Germination rate value of Pseudomonas fluorescens dose
treatment 100 ml l-1 significantly
different from the control treatment, and at the dose of Pseudomonas
fluorescens 200 ml l-1 which
has a germination rate value of 50.33% and 49.33% respectively. The dose of Pseudomonas
fluorescens was able to increase germination rate values by 12.67%
compared to control.
Pseudomonas fluorescens dose treatment200
ml l-1 produces the smallest germination rate value, not
significantly different from the control. Pseudomonas fluorescens dosage
200 ml l-1 had a negative impact on
germination rate values, reduced germination rate values by 1% compared to
control germination rate and was suspected of having a toxic effect on the seed
germination process. The relationship between Pseudomonas fluorescens dose
treatment and jatropha seed germination could be seen in Figure 11.
Figure 11. Effect of Pseudomonas fluorescens treatment
on jatropha seed germination
The results of orthogonal polynomial test produced a
regression equation (y) = -x2 + x + 50.333 with a coefficient of determination
(R²) = 1. Based on the regression equation it showed the optimal value at the
treatment dose of Pseudomonas fluorescens of 99.35 ml l-1 produces an optimal GC value of
63.16%. Pseudomonas fluorescens 100 dose ml l-1
in the exact treatment was the best dose in influencing jatropha seed
germination compared to other doses. Dose 100 ml l-1
Pseudomonas fluorescens was closed to the time that produces the optimal value of jatropha
seed germination.
Several studies have stated that matriconditioning was a combination of
giving Pseudomonas fluorescens density 109
cfu/ml or equivalent to dose of 100 ml l-1
in this study, could increase seed viability. According to Sutariati et al.
(2014), administration of Pseudomonas fluorescens dose of 109 cfu/ml used in matriconditioning invigoration,
had an impact on increasing germination rate values by 15% compared to
matriconditioning without Pseudomonas fluorescens on upland rice seeds. Pseudomonas
fluorescens dosage 109 cfu/ml used in the
matriconditioning invigoration of chili seeds had an impact on increasing
germination by 20%.
Maximum Growth
Potential (PTM %)
Based on DMRT test, Pseudomonas fluorescens dose of 100 ml l-1 produced the highest maximum
growth potential (PTM), compared to other treatments with a PTM value of
64.83%. Maximum growth potential from Pseudomonas fluorescens dose of 100 ml l-1 significantly different from
the control treatment and at the dose of Pseudomonas fluorescens 200 ml l-1 which has a PTM value of
53.58% and 53.50% respectively. PTM value of Pseudomonas fluorescens dose
of 100 ml l-1 significantly
different with dose of 200 ml l-1 and
control. Pseudomonas fluorescens dose of 100
ml l-1 was able to increase PTM by 11.25% compared to control.
Pseudomonas fluorescens dose of 200 ml l-1
produced values that were not significantly different with the
control, and produced the smallest Kst value. Pseudomonas fluorescens dose
of 200 ml l-1 had a negative impact
on PTM values, reducing PTM values by 1% compared to controls. The increase
of PTM was affected by the germination rate of the number of normal sprouts and
the number of abnormal sprouts, the dose of Pseudomonas fluorescens 200 ml l-1 increased the number of abnormal
sprouts by 1.99%. The relationship between the dose of Pseudomonas
fluorescens and the maximum growth potential of Jatropha seeds could be
seen in Figure 12.
Figure 12. Effect
of Pseudomonas fluorescens treatment on maximum growth potential of
Jatropha seeds
The results of the orthogonal polynomial test produced a regression
equation (y) = -x2 + x + 53,583 with a coefficient of determination (R²) = 1.
Based on the regression equation, it showed that the optimal value at the
treatment dose of Pseudomonas fluorescens is 102.46 ml l-1 produced an optimal PTM value of 65.13%. Pseudomonas
fluorescens dose of 100 ml l-1
was the best dose in influencing the maximum growth potential of jatropha seeds
compared to the other doses. Dose of 100 ml l-1 Pseudomonas fluorescensclose to the
time that produces the optimal value of the maximum growth potential of
Jatropha seeds. Application of Pseudomonas fluorescens used in
matriconditioning invigoration had an effect of increasing PTM values by 8%
compared to matriconditioning without Pseudomonas fluorescens on upland
rice seeds (Sutariati et al., 2014).
Discussion
Interaction
between treatments of matriconditioning invigoration and application of the
biological agent Pseudomonas fluorescens significantly affected the
observed variables of growth speed, vigor index and T50 (Table 3). The interaction occuring in the
growth rate variable (Kct) could explain that the combination of matriconditioning
invigoration treatment and the application of the biological agent Pseudomonas
fluorescens increases the vigor of Jatropha seeds in the form of an
increase in the value of growth speed. The combination of 24 hours of
matriconditioning and a dose of 100 ml l-1 Pseudomonas
fluorescens was the recommended treatment for use with a Kct value of
8.76%/etmal. Increasing the value of Kct had a positive effect on increasing
the viability (GR/GC) of Jatropha seeds, the two observational variables had a
strong positive correlation. The increase in growth rate also occurred in the
combination of matriconditioning treatment of roasted husk matrix and Pseudomonas
fluorescens at a dose of 109 cfu ml-1 which had a positive
effect on upland rice seeds, increasing Kct
up to 30 %/etmal (Sutariati et al., 2021).
The
interaction that occurs in the vigor index variable (VI) could explain that
combination of matriconditioning invigoration and Pseudomonas
fluorescensincrease Jatropha seeds increased vigor index. The combination
of 24 hours of matriconditioning time and a dose of 100 ml l-1 Pseudomonas
fluorescens was the recommended treatment for use with the best VI value of
61.33%. Increasing the value of VI had a positive effect on increasing the
viability (germination rate) of Jatropha seeds, the two observational variables
have a strong positive correlation. The results of this study are in line with
research Sutariati et al. (2021) explained
that VI
upland rice seeds increased by 52% compared to
the control without matriconditioning and combination treatmentPseudomonas fluorescens.
Other sources mentioned that the combination of matriconditioning and Pseudomonas
fluorescens increased VI
in chili seeds up to 17.33% and up to 20% in chili seeds infected with C.
acutatum
(F. A. Permatasari et al., 2016).
The
interaction occuring in the variable T50 could explain that combination of
matriconditioning invigoration and application of Pseudomonas fluorescens
accelerate seed growth time by 50% (T50). The combination of 24 hours of
matriconditioning without administration of Pseudomonas fluorescens was
the recommended treatment for use in increasing T50 values. The relationship
between the T50 value in influencing the germination value had a very low
correlation. Combination matriconditioning and Pseudomonas
fluorescensalso showed significant results in solving postharvest physiological dormancy in upland
rice (after ripening). This conclusion was seen based on the T50 value as an
indicator of seed dormancy, the T50 value decreased compared to the control
(5.04 days) to (4.47 days) after the combination treatment matriconditioning for 48 hours and Pseudomonas
fluorescens dose of 109
cfu ml-1 (Sutariati et al., 2021).
The
variables of growth rate and germination had a unidirectional relationship with
a very high degree of closeness. Each increase in seed vigor was represented by
a growth rate of 1%/etmal would be followed by an increase in seed viability
represented by seed germination of 5.59%. The combination of 24 hours of matriconditioning
and dose of 100 ml l-1 of Pseudomonas fluorescens increased
Kct, followed by an increase in seed viability (GC).This combination treatment was also able to
increase the Kct value in chili seeds by 3.74%/etmal and by 3.94%/etmal in
chili seeds infected with C. acutatum disease compared to controls (F. A.
Permatasari et al., 2016).The correlation between Kct and GC
values also occurs in sesame seeds with a correlation value (0.88) in the
same direction and closely (Hartati, 2019).
The
vigor index variable on seed germination has a unidirectional relationship with
a very high level of closeness. Each increase in Jatropha seed vigor is
represented by a vigor index of 10%, followed by an increase in seed viability
represented by germination of 7.94%. The increase in Jatropha seed viability in
the form of germination has a close relationship with the vigor index value, so
that the high germination value could be known from the first count germination
performance. Increases and decreases in the vigor index are always followed by
increases and decreases in germination. This occurs in several different types
of seeds, in papaya (Rosyad et al., 2016), in rice
seeds (Zhulfikri, 2018), in
cocoa seeds (Rachma et al., 2018). These
results are in accordance with (Hartati, 2019).
The results of this study are in line with the opinion
of Sadjad et al. (1999), that seeds having a high growth rate have a high level
of vigor. Seed viability and vigor are a unit of seed quality which has the
same cycle graph, forming a sigmoid-shaped curve (Ilyas, 2012). This
curve means that the decrease in quality that occurs in seeds is preceded by a
decrease in vigor and is followed by seed viability (Justice and Bass, 1979).
The duration of matriconditioning invigoration
significantly affected on growth rate (Kct), vigor index (VI), T50, growth
simultaneity (Kst) and germination capacity (GC). However, it had no
significant effect on the plant height, maximum growth potential (MGP), normal
sprout dry weight (NSDW) and abnormal seeds.
Based on the research result, the 24-hours matriconditioning
treatment produced the best Kst value compared to other matriconditioning
treatments. It was supported by the results of the polynomial regression
equation test which produced the optimal value of Kst closed to the 24 hours
matriconditioning treatment. Increasing the value of Kst has a positive effect
on increasing the viability (GC) of Jatropha seeds. The matriconditioning
invigoration of seeds for each plant species requires a different time
depending on the type of seed, size, physical seed and chemical content of the
seed. This statement was proven by the matriconditioning treatment for 5 hours
on cocoa seeds on the roasted husk matrix, which was the right length of
treatment, capable of increase the
Kst value by 28.5% (Rachma et al.,
2018). Matriconditioning research on peanut seeds
resulted in an increase in Kst of 26.67% in the roasted husk matrix with the
best treatment time of 12 hours (Muazizah, 2019).The
negative impact of the matriconditioning treatment in this study occurred at
the 48 hours level resulting in a Kst value of 47.11%, which was followed by a
decrease in the value of seed germination.
The 24-hours matriconditioning treatment resulted in
the best germination value compared to other matriconditioning treatments. The
combination of these treatments was supported by the results of the polynomial
regression equation test which produced the optimal GC value close to the 24
hours matriconditioning treatment. Increasing the vigor of jatropha seeds in
the form of Kct, vigor index, T50, MGP and Kst, had a positive effect on
increasing GC values. Invigorating seed matriconditioning for each plant
species required different time to increase seed viability and vigor, depending
on the type of seed, size, physical seed and chemical content of the seed.
Matriconditioning treatment for 48 hours on upland rice seeds on the burnt husk
matrix was the right treatment time, able to increase GC value by 65% (Sutariati et
al., 2021). The germination capacity of peanuts could be
increased to 26.67% through the matriconditioning invigoration treatment with
the roasted husk matrix for 12 hours (Muazizah, 2019).
The negative impact of the matriconditioning treatment
occurred at the 48 hour level resulting in a GC value of 47.11%, which was
followed by a decrease in the value of seed germination. A significant increase
in germination value is an indicator that 24 hours of matriconditioning time
could increase the percentage of seed viability. Negative impact of treatment matriconditioning on seed viability could occur due to the length
of treatment on the seed. Germination decreased by 2% in the matriconditioning
treatment of soybean seeds for 18 hours compared to no matriconditioning
treatment while the optimal length of treatment was 12 hours (Mariani & Wahditiya, 2021).
Matriconditioning treatment of purple egg plant seeds for 7 days could reduce
germination by 15.4%, lower than the best treatment time (Hasan et al.,
2018).
Application of Pseudomonas fluorescens
significantly affected on growth rate (Kct), vigor index (VI), T50, growth
simultaneity (Kst) and germination rate (GC). In contrast, it had no significant
effect on the plant height, maximum growth potential (MGP), normal sprout dry
weight (NSDW) and abnormal seeds.
Based on research results, Pseudomonas fluorescens 100
ml l-1 treatment produced the highest and best Kst value compared to
other treatments, by 63%. The effect of application of this dose of Pseudomonas
fluorescens increased the Kst of Jatropha seeds by 12.67% compared to the
Kst of seeds without Pseudomonas fluorescens application. It was
supported by the results of the polynomial regression equation test which
yielded an optimal value of Kst closed to the 100 ml l-1 dose of Pseudomonas
fluorescens. Increasing the value of Kst had a positive impact on
increasing the viability (GC) of Jatropha seeds. The need for biological agents
in seed treatment needed to be adjusted because they could have positive and
negative effects at certain doses on seed vigor and viability. The results of
this study were in line with research’s Sutariati
et al. (2021), the simultaneous growth of upland rice seeds
increased by 19% in the addition of treatment Pseudomonas
fluorescens density 109
cfu ml-1 compared to
matriconditioning without applicationPseudomonas
fluorescens.
The treatment of Pseudomonas fluorescens 100 ml
l-1 increased the percentage of germination and maximum growth
potential of Jatropha seeds. The highest and best germination percentage
achieved by 63%. This treatment increased the GC value of Jatropha seeds by
12.67% compared to control. It also was supported by the results of the
polynomial regression equation test which yielded the optimal GC value close to
the 100 ml l-1 dose of Pseudomonas fluorescens. The increase
in the GC value followed the increase in the vigor value of the seeds, the Kct
value and vigor index in this study had a positive and very close correlation
with the viability (GC) value of Jatropha curcas seeds. This research was in
line with the exposure of Sutariati et
al. (2021), treatment of Pseudomonas
fluorescens dose of 109 cfu ml-1 on capable upland rice seeds increase GC value 15%.
The treatment of Pseudomonas fluorescens 100 ml
l-1 could produce the highest maximum growth potential and achieve
the best by 64.83%,
an increase of 11.25% compared to control.
This combination of treatments was supported by the results of the polynomial
regression equation test which yielded optimal PTM values close to the 100 ml
l-1 dose of Pseudomonas fluorescens. The increase in PTM
value was determined by the increase in DB and the percentage of abnormal
seeds. The increase in the viability of Jatropha seeds in this study had a
positive impact on increasing the maximum growth potential of the seeds. The
positive impact on this treatment also occurred in the study of Sutariati et
al. (2021),
maximum growth potential upland
rice seeds increase 8% after
treatment Pseudomonas
fluorescens 109 cfu ml l-1 compared to the control.
The matriconditioning
treatment in this study increased viability in the form of germination capacity
(GC) compared to before treatment, vigor in the form of growth speed (Kct,
vigor index, T50) and growth simultaneity (Kst) in Jatropha curcas seeds. Seeds
that have been given matriconditioning treatment according to Sucahyono (2013) had metabolic conditions that were ready to
germinate, but the appearance of the radicle could be delayed so that the
percentage of germination and growth rate of sprouts increases when planted.
The process of seed germination occured through several stages,
matriconditioning could affect the mechanism and performance of jatropha seed
germination. Utilization of Pf also had the potential to prevent disease
attacks on plants (Harmaningrum, no year).
The factors influencing success matriconditioning according to Copeland and McDonald (2001), namely
conditions during matriconditioning (temperature and light), type and matrix
osmotic potential, oxygen availability, length of treatment time, control of
pathogen contamination, and method of seed drying. The matriconditioning
treatment of Jatropha seeds with the right length of time affected the success
of the aim of increasing seed viability and vigor. Pathogen control in the form
of matriconditioning integration with the biological agent Pseudomonas
fluorescens also influences the factors of increasing the viability and
vigor of Jatropha seeds.
Seed germination process according to Copeland and McDonald (2001),
consisted of several stages, starting with imbibition through the testa of the
seed, hydration of the aleurone of the seed up to the endosperm and embryo. The
next step was the activation of enzymes in seeds such as amylase, lipase and
protease enzymes in seeds. The enzyme in the next stage breaks down the food
reserves in the seeds in the form of proteins, fats, carbohydrates and others
into soluble forms. The soluble energy reserves were translocated to the
growing point of the embryo. The hormones auxin, gibberellins and cytokinins
worked to support the allocation of available energy to the area of cell
growth. The radicle and plumule grew through the seed coat, as a result of cell
growth, the process of cell enlargement, and the development of the sprout's
growing point.
Imbibition mechanism in
seeds given invigoration treatment matriconditioning
different from the process of seed germination in general without
treatment. Controlled imbibition in the matriconditioning treatment extended
the time for water absorption which allowed for the improvement of the
metabolic activity of Jatropha seeds. The imbibition mechanism provided controlled hydration to seeds which was controlled
by the physical strength of the moist solid media used (fired husks), had a low
matrix potential and negligible osmotic potential (Khan, 1992). The media used
according to Taylor et al. (1988), must had conditions that had water holding
capacity, be able to balance and release water according to the moisture
content of the seeds, not be toxic to the seeds and could be cleaned from the
seeds.
Seeds that experienced deterioration
experienced damage to the permeability and integrity of the cell membrane as
indicated by cell leakage on the results of the electrical conductivity test.
Free radicals caused damage to cell membranes that interfere with electron
transport, scatter free electrons which join with free radicals resulting in
damage to the integrity of germ cell membranes (Copeland and Mcdonald, 2001). Direct and
uncontrolled hydration could cause additional damage to cell membranes, because
seeds with low water content were prone to imbibition injury. A
controlled imbibition process would improve germination performance so that
seed viability and vigor could be improved, especially in seeds that experience
deterioration (Finch-Savage et al., 2004).Orthodox
seeds that were sensitive to cold conditions and high in fat, such as nuts,
were more susceptible to imbibition injury (Taylor et al., 1988).
Invigoration of matriconditioning
showed different results depending on the type of plant seed, the permeability of each type of
plant seed was different depending on the physical testa of the seed (thick,
hard, thin, soft), and the content in the seed (fat, protein, starch) would
affect the rate of imbibition (Finch-Savage et al., 2004). Orthodox seeds with
thick skin and high fat content, such as Jatropha curcas, were thought to
experience a slower imbibition process than other types of seeds. Hard testa
and high fat content in seeds that had low permeability properties slow down
the imbibition process, in some seeds it could be increased by
matriconditioning treatments (Taylor et al., 1988). Invigoration of
matriconditioning in some seeds found an improvement in the integrity of the
cell membrane, judging by the decrease in the resulting electrical conductivity
(EC).
The matriconditioning invigoration mechanism after imbibition helped
the activation of hormones and enzymes in Jatropha seeds last longer, so that
the internal potential of the seeds could be maximized. Enzymes that work in
the overhaul of seed food reserved during matriconditioning were amylase,
lipase and protease enzymes. The endogenous enzyme amylase in the seeds helped
break down food reserves in the form of starch into sugar (sucrose) as energy.
The lipase enzyme changed the lipids contained in Jatropha seeds into fatty
acids and glycerol and is broken down more simply into glucose and ends up as
energy. The protease enzymes in the seeds changed the protein preparations in
the seeds into amino acids and amides in forming new proteins for germination
(McDonald and Copeland, 1995). It happened during the time of matriconditioning
invigoration.
The performance of hormones in seeds when active
matriconditioning affected seed germination in the form of gibberellin (GA3),
auxin (IAA), and cytokinin hormones found in seeds. Cytokinins affected the
action of phytochromes, could change the permeability of cell membranes,
thereby allowing the release of gibberellin from the scutellum to the aleurone
during seed germination (Copeland et al., 2001). Cytokinins studied
in many studies affected cell division in germination and helped break seed
dormancy (Bradford and Nonogaki, 2007). Gibberellins influence seed germination
in mobilizing energy in seeds (Taiz and Zeiger, 2020), and in some
circumstances could replace light and temperature in germination (Copeland et al., 2001). The hormone of
auxin influenced embryo development and initiated the development of the basal
axis in seeds, namely the formation of plumule and radicle into apical shoots
and roots (Taiz and Zeiger, 2020). Auxin interacts with light to affect seed
germination, at high concentrations, it could inhibit germination (Copeland et al., 2001).
Influence of matriconditioning had been proven to
improve the vigor and viability of seeds studied from the metabolic activity of
the seeds, when the matriconditioning treatment supported the germination
process. Pathogen control as a factor influencing the success of
matriconditioning could be integrated with the use of beneficial biological
agents for increasing seed viability and vigor. The combination of
matriconditioning with beneficial microbes was a research that needed to be
further developed, the use of Pseudomonas fluorescens had been used with
excess production of enzymes and the resulting siderophores (Khan, 1992).
Administration of biological agents of Pseudomonas fluorescens influenced on
the viability and vigor of Jatropha seeds. The given effect had positive and
negative impacts depending on the concentration and dose of the biological
agent population. A positive effect was shown by an increase in the value of
viability (germination) and vigor (speed of growth, uniformity of growth, vigor
index) of Jatropha seeds at the dose of the biological agent of Pseudomonas
fluorescens 100 ml l-1. The increase in the viability of
Jatropha seeds was thought to have occurred due to the presence of hormones
from the secondary metabolites of the biological agent Pseudomonas
fluorescens. This statement was in accordance with the explanation from
Soesanto (2017), which
explains that these biological agents produced secondary metabolites in the
form of hormones, antibiotics, siderophores, enzymes and fats.
Biological agents Pseudomonas
fluorescens strain producing high cytokinins which was
propagated at 28ºC with a long shaker time of 72 hours, using NB (Nutrient
Broth) media capable of producing cytokinin hormones reaching 280 ppm (Kapoor
and Kaur, 2016). Cytokinin hormones played a role in the process of seed
germination, research by Un et al. (2018), explained that the hormone
gibberellins had an effect on increasing the viability of sandalwood seeds by
84%, breaking dormancy and increasing vigor reaching 2.4%/etmal. Cytokinin
hormones in this study could penetrate Jatropha seeds with thick skin types.
The mechanism of cytokinin hormones in the germination process was to encourage
cell division and enlargement, stimulate the embryo and cotyledons to give rise
to the seed plumule,
Biological agents of Pseudomonas fluorescens produced secondary metabolites, in the form
of the hormone auxin indole acetic acid (IAA) (Istiqomah et al., 2017). the
bacteria could synthesize IAA up to 68.9 ppm (Tefa, 2018). The auxin hormone produced depends on the
nutrients used and the incubation growing environment in the multiplication of
these biological agents. The IAA hormone had a positive effect on increasing
mung bean germination up to 24% and spurring mung bean root elongation (Janani
et al., 2017). This hormone was thought to cause an increase in Jatropha seed
vigor in the form of an increase in the value of growth speed, vigor index and
the value of growing simultaneity in sprouts. The IAA hormone produced by
biological agents played a role in influencing the physiology of seeds,
stimulating the growth of the radicle and plumule of the seed during the
germination process (Herlina et al., 2017). The auxin mechanism in seed
germination started with the hormone penetrating the testa of the seed,
entering the embryo and cotyledons,
Hormones were produced by the biological agent
of Pseudomonas fluorescens apart from the auxin and cytokinin types was
also capable of producing hormones of gibberellins reach 69.2 ppm (Tefa, 2015),
25-60 ppm (Kapoor et al., 2016). Gibberellin hormone in the study of Un et al.
(2018), explained that it had an effect on increasing the viability of
sandalwood seeds by 84%, breaking dormancy and increasing vigor reaching
2.4%/etmal. Hormones produced by the biological agent Pseudomonas
fluorescens are thought to influence seedling growth and affect germination
speed, growth synchrony and the number of Jatropha sprouts. The hormone
gibberellin played a role in influencing seed germination in mobilizing the
available food reserves in the seed, so that it begins to be broken down into
energy, weakening the seed coat that caused dormancy (Taiz and Zeiger, 2020).
Food reserves stored in jatropha seeds in the form of
protein, fat, starch, phosphate were used by the seeds as energy and carbon
sources after being hydrolyzed by hydrolytic enzymes in the seeds or from
exogenous products produced by biological agents. The enzymes produced by the
biological agent Pseudomonas fluorescens were protease enzymes,
chitinase enzymes, cellulases, lipases and amylase enzymes (Soesanto, 2017). Each enzyme
produced by these biological agents had a different use, and was thought to
influence the process of seed germination. Jatropha seeds that had been imbibed
penetrate the seed testa with a soluble amylase enzyme, which functions to
catalyze starch in the seed which was converted into soluble sugar in the form
of maltose. The dissolved sugar was used as energy for the seed embryo to
germinate.
The protease enzyme produced by the biological agent Pseudomonas
fluorescens catalyzes the protein contained in Jatropha seeds. Proteins
that had undergone hydrolysis into amino acids become proteins in a form
available to seed embryos in the germination process (Joshi, 2018). Kawi
Researchnski et al. (2021), explained that the lipase enzyme
in seeds was shown to change the amount of hydrolyzed fat into free fatty acids
and glycerol during the germination process of jojoba seeds. According to
Sugiharni (2010), the lipase enzyme produced by Pseudomonas fluorescens had
been shown to have a function as an enzyme that could hydrolyze fats produced
by plant seeds into soluble fatty acids and glycerol which seeds use for
germination.
The biological agent of Pseudomonas fluorescens in
seed treatment was useful as a bioprotectant presumably due to compounds
produced from secondary metabolites in the form of antibiotics and
siderophores. Antibiotics produced in this study could harm and inhibit the
development of other microbes. The antibiotics produced by these biological
agents were salicylic acid, pseudomonic acid, cyanhydrate,
2,4-diacetylfluorolucinol (Soesanto, 2017). Antibiotic
compounds produced by biological agents could inhibit the growth of pathogens
that attack seeds and plants. Permatasari (2016) showed that the
results of the Pseudomonas fluorescens antagonist test could inhibit the
growth of C. acutatum, followed by the same results in field conditions
on chili plants. Apart from antibiotics, Pseudomonas fluorescens also
produced siderophore compounds which were useful as inhibitors of pathogen
growth. Siderophores were special compounds produced by Pseudomonas
fluorescens which were useful for binding iron in the growth environment,
so that they could suppress the growth of pathogens (Soesanto, 2017).
Pathogenic attack still
occured in some sprouts. Sprouts attacked by pathogens had
characteristic rot on the stem but did not attack the roots. The pathogen from
the sprouts was grown on agar media and it was seen that the disease originated
from a fungus with characteristic white hyphae which turned brown after one
week of incubation. Even though it was not observed microscopically, it was
found that the cause of the stem rot of the sprouts came from a fungus. These
pathogens could come from Jatropha seeds (seed-borne diseases) or from the
growing environment. Seeds that were damaged by the testa, were opened and
inside there were brown hyphae. Washing with 5% sodium hypochlorite during 5 minutes and rinsed with distilled water, to
reduce the incidence of diseases that may be caused by seed-borne pathogens (Sutariati et
al., 2021).
CONCLUSION
Based on the results and discussion above it could be concluded
that; (1) the
interaction effect of matriconditioning time and dose of Pseudomonas
fluorescens was significantly different on the observed variables of growth
speed, vigor index and T50. Combination of 24 hours matriconditioning treatment
and dose of Pseudomonas fluorescens 100 ml l-1
produced the best treatment on variables representing seed vigor (growth rate and vigor index),
and (2) the effect of a single factor on
matriconditioning time was significantly different and the best was the 24-hour
treatment period seed viability (germination) and
seed vigor (growing rate,
growth synchrony and vigor index).The effect of a single factor dose of Pseudomonas
fluorescens was significantly different and the best at the dose 100
ml l-1 to seed viability (germination), seed
vigor (growth rate,
growth simultaneity, vigor index) and maximum growth potential.
REFERENCES
Afa, L. O. (2008). Peningkatan Viabilitas Benih Jati (Tectona grandis Lf)
dengan Tehnik Invigorasi Benih Menggunakan Biomatriconditioning Pseudomonas
fluorescens. AGRIPLUS18, 3, 187–194.
Copeland, L. O., McDonald, M. B., Copeland, L. O.,
& McDonald, M. B. (2001). Seed germination. Principles of Seed Science
and Technology, 72–123.
Hartati, P. (2019). Hubungan Deteriorasi dengan Umur
Simpan Benih Melalui Penggunaan Indiktor Pengujian Viabilitas dan Vigor pada
Benih Wijen (Sesamum indicum L.). Universitas Sumatera Utara.
Hasan, A., Taufik, M., Kasim, N. N., & Hijria, H.
(2018). Deteksi Soybean mosaic virus (SMV) terbawa benih kedelai di Sulawesi
Tenggara. Journal TABARO Agriculture Science, 2(1), 201–205.
Hasanuddin. (2010). Effect of Maturity Level and Storage
Period on Jatropha Seed Viability (Jatropha curcas L.). Bogor Agricultural
Institute.
Ilyas, S. (2012). Ilmu dan teknologi benih: Teori
dan hasil penelitian. PT Penerbit IPB Press.
ISTA. (2014). International rules for seed testing.
Istiqomah, I., Aini, L. Q., & Abadi, A. L. (2017).
Kemampuan Bacillus subtilis dan Pseudomonas fluorescens dalam melarutkan fosfat
dan memproduksi hormon IAA (Indole Acetic Acid) untuk meningkatkan pertumbuhan
tanaman tomat. Buana Sains, 17(1), 75–84.
Kapoor, R., & Kaur, M. (2016). Cytokinins
production by fluorescent Pseudomonas isolated from rhizospheric soils of Malus
and Pyrus. African Journal of Microbiology Research, 10(32),
1274–1279.
Khan, A. A. (1992). Preplant physiological seed
conditioning. Horticultural Reviews, 13(1), 131–181.
Mariani, M., & Wahditiya, A. A. (2021). Pengaruh
Perlakuan Matriconditioning Terhadap Viabilitas Dan Vigor Benih Kedelai
(Glycine max L. Merrill). Jurnal Agrotan, 7(1), 55–67.
Muazizah, T. (2019). Perlakuan Invigorasi Terhadap
Viabilitas Beberapa Varietas Benih Kacang Tanah (Arachis hypogaea L.).
Universitas Sumatera Utara.
Permatasari, F. A., Aimon, A. H., Iskandar, F., Ogi,
T., & Okuyama, K. (2016). Role of C–N configurations in the
photoluminescence of graphene quantum dots synthesized by a hydrothermal route.
Scientific Reports, 6(1), 1–8.
Permatasari, O. S. I., Widajati, E., & Syukur, M.
(2016). Aplikasi bakteri probiotik Pseudomonas kelompok fluorescens untuk
meningkatkan produksi dan mutu benih cabai. Jurnal Agronomi Indonesia (Indonesian
Journal of Agronomy), 44(3), 292–298.
Priyanto, Y. A. (2017). (Viabilitas Benih Kedelai
(Glycine Max. L. Merril) dengan Perlakuan Invigorasi Matriconditioning dan
Osmoconditioning). Jurnal Hexagro, 1(1), 292637.
Rachma, T. N. S., Damanhuri, D., & Saptadi, D.
(2018). Viabilitas dan Vigor Benih Kakao (Theobroma cacao L.) pada Beberapa
Jenis Media Invigorasi. Plantropica: Journal of Agricultural Science, 1(2).
Rosyad, A., Suhartanto, M. R., & Qadir, A. (2016).
Pola Penurunan Viabilitas dan Pengembangan Metode Pendugaan Vigor Daya Simpan
Benih Pepaya (Carica Papaya L.). Jurnal Hortikultura Indonesia, 7(3),
146–154.
Rustam, R., & Audina, M. (2018). Uji Tepung Biji
Mengkudu (Morinda Citrifolia L.) Terhadap Hama Bubuk Jagung Sitophilus Zeamais
M.(Coleoptera; Curculionidae). Jurnal Agroekoteknologi, 10(1).
Sadjad, S. (1989). Konsepsi Steinbauer-Sadjad sebagai
landasan pengembangan matematika benih di Indonesia. Pidato Ilmiah
Pengukuhan Guru Besar. Fakultas Pertanian, Institut Pertanian Bogor. Bogor.
Scales, B. S., Dickson, R. P., LiPuma, J. J., &
Huffnagle, G. B. (2014). Microbiology, genomics, and clinical significance of
the Pseudomonas fluorescens species complex, an unappreciated colonizer of
humans. Clinical Microbiology Reviews, 27(4), 927–948.
Soesanto, L. (2017). Pengantar Pestisida Hayati Adendum
Metabolit Sekunder Agensia Hayati. Rajawali Pers. Jakarta.
Sucahyono, D. (2013). Invigorasi benih kedelai. Buletin
Palawija, 25(2), 18–25.
Sutariati, G. A. K., Khaeruni, A., Madiki, A., Mudi,
L., Aco, A., Ramadhani, S. A., Adri, R. M., & Mantja, K. (2021).
Effectiveness of endo-rhizobacteria as growth promoters and biological control
of moler disease in shallots (Allium ascalonicum L.). IOP Conference Series:
Earth and Environmental Science, 681(1), 12028.
Sutopo, L. (2010). Teknologi Benih (Edisi Revisi
Fakultas Pertanian UNIBRAW). PT Raja Grafindo Persada.
Tefa, A. (2015). Pemanfaatan bakteri probiotik untuk
menekan infeksi Colletotrichum acutatum dan meningkatkan mutu benih cabai
(Capsicum annuum L.) selama penyimpanan. Tesis]. Sekolah Pascasarjana
Institut Pertanian Bogor. Bogor.
Tefa, A. (2018). Perlakuan Invigorasi Pada Benih Padi
di Kelompok Tani Pelita Desa Noepesu. Bakti Cendana, 1(1), 1–10.
Yuniarti, N. (2020). The Invigoration Techniques of
Nyamplung (Calophyllum inophyllum L.) Seeds During the Storage. Jurnal
Wasian, 7(1), 65–72.
Yunita, R., Krisyanella, K., Pudiarifanti, N.,
Iqoranny, A., & Irnameria, D. (2021). Uji Aktivitas Antibakteri Ekstrak
Daun Sawo (Manilkara zapota L) Terhadap Bakteri Pseudomonas aeruginosa.
Poltekkes Kemenkes Bengkulu.
Zanzibar, M. (2016). Rapid estimation of the
viability of forest plant seeds, principles of methods and their applications.
Self-help spreader.
Zhulfikri, M. (2018). Penentuan Waktu Pengujian
Cepat Vigor Benih Padi IPB 3S dengan Metode Radicle Emergence.
Copyright holders:
Sundahri, Tofan Mursyidto, Tri
C. Setiawati, Hardian A. Susilo, Ali Wafa (2023)
First publication right:
Devotion - Journal of Research and Community
Service
This
article is licensed under a Creative
Commons Attribution-ShareAlike 4.0 International