Effect of moisture level and storage container on the

Bull. Inst. Trop. Agr., Kyushu Univ. 36: 61-69, 2013

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Effect of moisture level and storage container on the quality of chickpea seed (Cicer arietinum) M. R. Islam1), M. A. Rahman2), M. M. Rashid3) and M. Shahin-Uz-Zaman4)

Abstract The effect of moisture levels and storage containers on the seed quality of chickpea was investigated in the laboratory of Department of Agronomy, Hajee Mohammad Danesh Science and Technology University (HSTU) Dinajpur, Bangladesh during April-July 2009. Three seed container viz, sealed container, polythene bag and gunny bag and three moisture level i.e. 8.88 %, 12.23 % and 14.10 % were included in the study. Chickpea seeds with 8.88% initial moisture level were found in good condition in respect of germination, less number of abnormal and dead seedlings, better shoot and root length, vigour index and poor fungal incidence. The highest germination percent was found 81.12−88.36%, while the lowest percent of abnormal seedling (9.21−13.00%) and dead seedling (3.39−7.20%) was found in chickpea with 8.88% the initial moisture level at different storage periods. Fungal incidence was found less in chickpea seeds with the same initial moisture level. Among the storage condition, sealed container was found the advantage for chickpea seed storage compared to the usage of polythene bag and gunny bag. The seed container of gunny bag was also found in inferior condition in case of above parameters. Interaction between sealed container and moisture level at 8.88% was performed better in all aspects compared to the gunny bag with 14.10% moisture level. Our results concluded that optimum moisture level and ideal storage condition has the greatest benefit on the quality seeds.

Keywords: Moisture levels, storage containers, storage periods, seed quality, chickpea

Introduction Deterioration of seed quality is more liable to humid sub-tropical climate. In Bangladesh, relative humidity is detrimental to seed preservation. From middle to east side of this country, it is having mean annual rainfall 2000-5000 mm and western part having average rainfall 1400-2000 mm., respectively (Ahmed, 1988). High rainfall cause high relative humidity and resulting in the high moisture level of seed. By staying in this condition, the seeds lose germination ability with low vigor seedlings at the end of storage. As a result, the seed quality becomes to inferior and the seed is unusable for crop production. So, seeds are not safe in our country for long time preservation. In our country, storage period of almost 1) Scientific officer, Agronomy Division, Regional Agricultural Research Station, BARI, Ishurdi, Pabna, Bangladesh 2) Professor, Department of Agronomy, Hajee Mohammad Danesh Science and Technology University (HSTU), Dinajpur, Bangladesh 3) Professor, Department of Plant pathology, Hajee Mohammad Danesh Science and Technology University (HSTU), Dinajpur, Bangladesh 4) Scientific officer, Pulses Breeder, Pulses Research Centre (PRC), BARI, Ishurdi, Pabna, Bangladesh

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all kinds of seeds is April to October. But unfortunately high rainfall, relative humidity and temperature prevail in this period cause detrimental effect to the seed longevity. Seed quality is the most important input in agricultural production. It ensures better germination as well as better yield. But the major hindrances to increasing seed production are the loss of seed viability and damage of seeds by biotic and abiotic factors during storage period. Majority of the farmers in Bangladesh, seeds are stored for three to six month before consumption. Traditionally, the farmer stored the seeds in earthen vessels (Motka), bamboo containers (Dole), jute sack or plastic bags. However, these were not suitable to protect the seed from the microbial infestation and can easily penetrate into the containers and infest to the stored seed. So, appropriate container can save the seed from deterioration of seed quality and viability. Moreover, as seed is highly hygroscopic living material; it absorbs moisture from air if it is stored in an environment where relative humidity is higher than seed moisture level (Copeland, 1976). Therefore, storage containers are most important for storing. Poor storage conditions greatly affect seed vigor (Heydecker, 1979). Seed longevity decreased with increase of storage temperature and moisture level and the deterioration rate depends on the storage condition which are including of temperature, relative humidity, seed moisture levels and storage container (Usberti et al., 1998). Types of container also regulate the condition of temperature, relative humidity and moisture levels of seed. The seed moisture level, their location and availability are the most important factors for influencing microbial growth. Almost of all microorganisms require the moisture for growth and growing rapidly in the presence of plentiful supply of moisture (Frazier, 1968). Most of the storage pathogenic species are Aspergillus spp, Penicillium spp and Rhizopus spp. which cause discoloration of the seeds and germination failure (84.4%) and yield loss (60%) development of the symptoms having color rot (84.4%) and wilt (60%), respectively (Fakir,1983). Aspergillus flavus, Aspergillus niger, Fusarium spp. and Penicillum oxalicum are common fungi for infection of chickpea seeds (Dwivedi, 1989). Good seed alone can give an increased production (10 to 15%) compared to the poor seed stock (Alim, 1977). So, storage of seed is an important factor for maintaining their seed quality. However, proper control of seed moisture level and seed storage conditions is essential for the maintenance of seed viability and vigor, in the humid tropical regions. In view of above factors, the present research work was undertaken to know the effect of moisture levels and storage containers on the seed quality of chickpea.

Materials and Methods The experiment was conducted in the Laboratory of Department of Agronomy Division, Hajee Mohammad Danesh Science and Technology University (HSTU), Dinajpur, Bangladesh, during April− July 2009 to investigate the effect of moisture levels and storage containers on the seed quality of chickpea. The experiment was laid out in a Completely Randomized design (CRD) with three replications. Three seed containers (sealed container, polythene bag (Thickness 0.05mm) and gunny bag) and three seed moisture levels (8.88%, 12.23% and 14.10 %) were included in the study. Fully matured chickpea seeds were (14.98% moisture level) collected from the local market, equilibrated to moisture levels (14.10%, 12.23% and 8.88%) by sun drying and stored for two month in sealed container, polythene bag

Effect of moisture level and storage container of chickpea seed

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and gunny bag at room temperature with relative humidity. The sealed pot was covered tightly; the polythene bag and gunny bag were tightly with rope. The initial germination percentage was 92.25%. During the storage period, hundred seeds samples were taken at every 20 days interval from the each container for determination of the germination percentage, abnormal and dead seedling, shoot and root length, vigour index and fungal incidence. Moisture level was determined by high constant temperature oven method following international rules for seed testing (Anonymous, 2000). Three grams of seeds from each container were used. After grinding the seeds in grinding mill, the weighed ground materials were poured separately in a small container with cover and kept in an oven maintained at a temperature of 125-130˚C for 2 h. The moisture level of seeds (wet basis) was determined by the following formula (Anonymous, 2000). %MC =

(X2 – X3) x100 (X2 – X1)

X1 = Weight of container X2 = Weight of container + ground materials before drying X3 = Weight of container + ground materials after drying Germination test was conducted using a sand as the substratum. The sand was sieved to discard particles bigger than 0.8 mm and smaller than 0.05 mm in diameter. Rectangular plastic boxes were used to put the new sand for every test. Seed was placed on a uniform layer on moist sand and then covered to a depth of 10 mm with sand, which was left loose. One hundred seeds were planted in each plastic tray and replicated for three times. The plastic trays with seed were incubated at room temperature and irrigated at every odd day. Eight days after sowing, the seed germination percentage was recorded. The normal seedlings, abnormal seedlings and dead seedling were classified according to the prescribed rules given by ISTA (1996). Germination (%) =

Number of normal seeds germinated x100 Number of seeds tested

For vigour index or seedling vigour data was recorded on the germination up to 10 days of sowing. Then root length and shoot length were measured from the seedling of pot for calculating vigour index. Vigour index (VI) was calculated by the formula reported by Baki and Anderson (1973) as shown below: Vigour index (VI) = Germination (%) × (Mean shoot length + mean root length) To detect the seed borne pathogens associated with the seeds in samples, the blotter method was used by following international rules for seed testing (ISTA 1996). In this method, three layer of blotting paper were soaked in sterilized water, placed at the bottom of 9 cm diameter plastic petridish and then 10 seeds were planted. The seeds in the petridish were incubated at 20±2˚C under alternating cycle of 12 hours near ultraviolet (NUV) light at darkness for 8 days. Time to time watering was maintained with the filter paper moist. Each individual incubated seed was observed under stereomicroscope at the 16x and 25x magnifications in order to record the incidence of seed borne fungi. Most of the associated pathogens were detected by observing their morphological traits on the incubated seeds on blotter paper. For proper identification of fungi, temporary slides were prepared from the fungal colony and observed

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with compound microscope and identified following the key outlined by Booth (1971) and Singh (1982). During the storage period, monthly average of maximum and minimum temperature ranged from 32.2 − 34˚C and 21 − 24.3˚C, respectively. The average relative humidity ranged from 81 to 87%. Data on germination percentage, abnormal seedling, dead seedling, shoot and root length, vigor index and infestation of fungus was recorded and statistically analyzed. Moreover, mean values were adjusted by LSD at 0.05 level of probability.

Results and Discussion Effect of moisture level Germination percentage at the different storage period was presented in Fig. 1. Two months after storage, the germination of chickpea seeds at all the moisture levels ranged from 60.31% to 81.12%. The highest germination (88.36%) gave the moisture level at 8.88% followed by 12.23%, while the minimum germination (69.40%, 66.26% and 60.31% in 20, 40 and 60 days after storage (DAS), respectively) was observed from the chickpea seeds stored at 14.10% moisture level. The result also revealed that 8.88% moisture level was found good up to 2 month storage with the germination percentages at 81.12%, while the germination rate was recorded poor as 74.63% and 60.31% with moisture level at 12.23% and 14.10%, respectively. Delouche (1968) reported that seed should never be stored at moisture levels higher than 14.50%, because they loss viability at the faster rate compared to the lower moisture level. Higher moisture level in seed during storage is one of the main reasons for loses of viability sooner. The moisture accelerates the respiration rate of seed and microorganism. A higher moisture level may produce heat rapidly enough to deteriorate seed quality (Brandenburg et al., 1961). Moreover, high moisture initiates incomplete physiological process of seed germination resulting in loss of viability (Harrington, 1972). ML (8.88%)

100

ML (12.23%)

ML (14.10%)

Germination percentage

90 80 70 60 50 40 30 20 10 0 20

40

60

Days after storage Fig. 1. Germination percentage of chickpea seed stored in different moisture levels.


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The moisture level of 8.88% was found good result in respect of producing little number of abnormal and dead seedlings (Table 1.). Abnormal seedling of chickpea was significantly affected by the moisture level for all storage periods and decreased with the increase of storage period in a given moisture levels. The maximum abnormal seedlings was recorded in chickpea seeds with 14.10% moisture level and the lowest abnormal seedlings was recorded in 8.88% moisture level at 20, 40 and 60 DAS, respectively. Moisture level exhibited significant effect on the dead seedling of chickpea for all storage periods. Dead seedling was increased with the increased of storage period. The highest dead seedlings was recorded in 14.10% moisture level and the lowest dead seedling was recorded in 8.88% moisture level at 20, 40 and 60 DAS, respectively. The results indicated that 8.88% moisture level is suitable for storage of chickpea seeds. These results are in agreement with the finding of Afzal et al., (2008) who reported that 8-9% moisture level is suitable for storage of chickpea seeds. The amount of moisture in the seeds affected the seed viability during storage. It is interred that the increase rate of deterioration is agreed with the increase of moisture level in seed (Agrowal, 2003). The highest shoot and root length were observed in chickpea seeds with the moisture level of 8.88%. The lowest shoot and root length was also occurred in chickpea seeds with 14.10% moisture level at different storage periods (Table 2). Copland (1967) reported that seedling vigor (growth rate) decline rapidly with the length of seed storage. Vigour index or seedling vigour was significantly influenced by the moisture level for all storage periods (Table 3). Vigour index was decreased with the increase of moisture levels and storage period. The highest vigour index was obtained from 8.88% moisture level, which was followed by polythene bag and the lowest vigour index was recorded in 14.10% moisture level for all storage periods. Vigour index decreasing gradually upto 60 DAS more in 14.10% moisture level might be due to the low vigour of seeds for long storage periods and fungal infestation. Incidence (%) of Fusarium oxysporum at differTable 1. Effect of moisture level and storage container on the abnormal and dead seedling of chickpea stored seed at different storage period Treatment

ML (8.88%) ML (12.23%) ML (14.10%) LSD(0.05) CV (%) Sealed container Polythene Gunny bag LSD(0.05) CV (%) ML × SC

Abnormal seedling (%) at 20DAS 40DAS 60DAS Effect of moisture level 9.21 11.18 13.00 10.86 13.47 17.56 19.53 20.92 25.57 1.71 1.41 1.60 11.23 8.05 7.42 Effect of container 10.40 12.94 16.26 11.82 13.89 18.00 17.38 18.74 21.87 0.94 1.06 0.95 7.00 6.82 4.99 Interactions NS NS *

20DAS

Dead seedling (%) at 40DAS 60DAS

3.39 5.95 11.39 1.47 18.51

4.17 8.58 12.80 1.42 14.52

7.20 11.13 14.78 1.32 10.38

4.94 6.47 9.32 0.95 13.47

6.40 8.18 10.98 0.71 8.22

8.64 10.23 14.24 0.96 8.55

NS

NS

*

DAS = Days after storage, ML = Moisture level, SC = Storage container, NS = Not significant * = Significant at 5% level probability

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Table 2. Effect of moisture level and storage container on shoot and root length of chickpea stored seed at different storage period Treatment 20DAS ML (8.88%) ML (12.23%) ML (14.10%) LSD(0.05) CV (%)

18.41 15.54 11.66 1.39 7.97

Sealed container Polythene Gunny bag LSD(0.05) CV (%)

16.25 15.27 14.08 0.81 5.24

ML × SC

NS

Shoot length (cm) at 40DAS 60DAS Effect of moisture level 17.46 16.04 14.82 13.03 11.86 10.26 1.32 1.60 7.82 10.58 Effect of container 15.94 13.99 14.97 13.22 13.23 12.13 1.55 0.97 10.25 7.26 Interactions NS NS

20DAS

Root length (cm) at 40DAS 60DAS

16.65 14.18 9.56 1.33 8.60

15.38 13.34 9.14 1.16 7.96

14.79 12.23 8.71 1.64 11.99

14.18 13.51 12.70 0.98 7.15

13.56 12.60 11.70 0.79 6.10

12.86 11.97 10.90 1.31 10.75

NS

NS

NS

DAS = Days after storage, ML = Moisture level, SC = Storage container, NS = Not significant Table 3. Effect of moisture level and storage container on vigour index and incidence (%) of Fusarium oxysporum of chickpea Treatment

Vigour index at 20 DAS

ML (8.88%) ML (12.23%) ML (14.10%) LSD(0.05) CV (%)

3097.90 2491.72 1472.67 240.10 4.50

Sealed container Polythene Gunny bag LSD(0.05) CV (%)

2575.90 2348.16 2018.14 208.70 3.98

ML × SC

NS

40DAS 60DAS Effect of moisture level 2790.41 2500.93 2232.52 1885.15 1391.46 1144.08 172.30 258.10 3.55 6.18 Effect of container 2398.94 2051.88 2139.16 1824.51 1784.99 1547.85 146.80 189.80 3.07 4.63 Interactions NS NS

Incidence (%) of Fusarium oxysporum at 20 DAS 40DAS 60DAS 3.20 5.73 8.00 1.14 17.53

3.76 6.60 9.17 1.16 15.44

4.69 8.12 11.39 0.88 9.46

4.87 5.23 6.83 0.62 10.84

5.61 6.13 7.78 0.84 12.61

6.96 7.22 10.02 0.50 6.11

NS

NS

NS

DAS = Days after storage, ML = Moisture level, SC = Storage container, NS = Not significant

ent moisture level and storage periods is presented in Table 3. The Maximum incidence of F. oxysporum were recorded from the seeds of chickpea stored at 14.10% moisture level in 20, 40 and 60 DAS, respectively. The lowest incidence (%) of F. oxysporum was recorded in the seeds of 8.88% moisture level followed by the seeds of 12.23% and 14.10% moisture level. The aforesaid fungal species are noted for their destructive characteristics in storage causing seed rotting and germination failure (Christensen et al., 1965). The highest germination and lowest prevalence of fungi was recorded in the seeds stored in the sealed container fallowed by polythene bag and gunny bag (Rahman et al., 1997).


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Effect of Container Germination percentage of stored chickpea seeds in different containers were decreased with increase of the storage period (Fig. 2). The effect of storage containers on germination of chickpea seed was statistically significant for all storage periods. The highest germination was recorded for seeds in sealed container and the lowest germination was recorded in gunny bag at 20, 40 and 60 DAS, respectively. The germination rate in sealed container, polythene bag and gunny bag at 20 DAS was 84.65, 81.59 and 75.36%, respectively. After 2 month, it was declined to 76.42, 72.43 and 67.21%. The decreasing rate was higher in seeds of gunny bag. Storage of seeds in gunny bags and the other local storing environment may influence to the increase of moisture and deteriorate seed quality (Copland 1976). Germination is the most important function as an indicator of its viability and worth to the seed. Germination percentage was decreased gradually upto 60 DAS in Gunny bag might be due to high moisture and fungi infestation. Storage of chickpea seeds in different containers and duration of storage showed significant effect on the production of abnormal and dead seedling (Table 1.). The entire storage container found higher percentage of abnormal seedling with advancement of the storage period. However, the minimum percentage of abnormal seedling (10.40%) was found in the seeds that were stored in sealed container for 20 days. But the maximum (21.87%) was found from the seeds that were stored in gunny bag for 60 days. Mali et al. (1983) reported that increasing rate of abnormal seedling was higher in seeds of gunny bag, because it was due to the condition of high moisture and fungal activities. Significant difference on the dead seedling of chickpea was observed among the storage containers for all storage periods. Dead seedling was increased with the increase of the storage period. Gunny bag gave the highest dead seedling and sealed container gave the lowest dead seedling at 20, 40 and 60 DAS, respectively. This con-

Sealed container

90

Polythene bag

Gunny bag

Germination percentage

80 70 60 50 40 30 20 10 0 20

40

60

Days after storage Fig. 2. Germination percentage of chickpea seed stored in different containers.

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tainer type affects moisture absorbance and seed quality during storage of seeds. The rate of absorbance was higher in gunny bag. The gunny bag is not air tight but sealed container and polythene bag are moisture proof. Since, moisture increasing rate was lower in sealed container. Higher moisture in the seed is the main reason of quick deterioration of the seed quality of gunny bag. Hence, the seeds of gunny bag produce the large number of abnormal and dead seedlings. The combination of initial shoot and root length of seeds in sealed container (16.25 & 14.18cm), polythene bag (15.27 & 13.51cm) and gunny bag (14.08 & 12.70cm), respectively, were presented in Table 2. At the end of 2 months, the shoot and root length were decreased to 13.99 & 12.86cm, 13.22 & 11.97cm and 12.13 & 10.90cm, respectively. The decreasing rate was higher in seeds of gunny bag. It might be due to more absorbance of moisture from the surrounding atmosphere, creating the favorable condition for microorganism. Hence, the seedling vigor was poor in the seeds of gunny bag. Seed viability and vigor significantly depends on the type of storage container (Ali, 1963). Deterioration of seeds during storage can cause significant declines in seedling vigor and crop yield (Biabani et al., 2011). Vigour index or seedling vigour and incidence (%) of F. oxysporum of chickpea at different containers and storage periods are presented in Table 3. Storage containers had significant effect on the vigour index that was decreased with the increase of storage period. The highest vigour index (2575.90) was observed from sealed container which was significantly differed from polythene bag (2348.16) and gunny bag (2018.14) at 20 DAS. In case of 60 DAS, vigour index ranged from 1547.85 to 2051.88, while the highest and lowest vigour index were recorded in sealed container and gunny bag, respectively. Storage containers had significant effect on the incidence of F. oxysporum for all storage periods. Incidence of F. oxysporum was increased with the length of storage period. The highest incidence of F. oxysporum was observed in gunny bag and the lowest incidence was observed in sealed container which was followed by polythene bag at 20 and 60 DAS, respectively. These results are in agreed with the findings of Khalequzzaman et al., (2012) who reported the highest vigour index and lowest incidence (%) of F. oxysporum in Tin container and lowest vigour index and highest incidence (%) of F. oxysporum in gunny bag at french bean. Khalequzzaman et al., (2004) also found that 19.99% seeds were infected by Fusarium spp. and 19.65% seeds were infected by Rhizopus sp. in blotter method of french bean seed. Interaction effect The interaction effect between moisture levels and storage containers had significant effect on the abnormal and dead seedling at 60 DAS (Table 1). Among the others parameters showed not significant at different days after storage, however, better performance obtained from the seed store in the sealed container with 8.88% moisture level in all aspects compared to the seed store in gunny bag with 14.10% moisture level.

Conclusion In this experiment, we concluded that sealed container and moisture level at 8.88 % showed comparatively better performance than other container (polythene bag and gunny bag) and are also presented in moisture levels (12.23% and 14.10%) in case of germination, less number of abnormal and


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dead seedling, shoot and root length, vigour index and poor fungal incidence of chickpea seeds at the storage condition.

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