Sorghum (Sorghum bicolor (L.) Moench.) Seed Quality as Affected by Type and Duration of Storage

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1 AGRICULTURE AND BIOLOGY JOURNAL OF NORTH AMERICA ISSN Print: , ISSN Online: , Science Huβ, Sorghum (Sorghum bicolor (L.) Moench.) Seed Quality as Affected by Type and Duration of Storage Eltayeb Elhag Ali Ahmed and Sana Hassan Ahmed Alama Dept. of Botany and Agric Biotechnology, Faculty of Agriculture, University of Khartoum, Shambat 13314, Sudan, Corresponding author: Eltayeb Elhag Ali Ahmed, phone number , ABSTRACT The objective of this study was to investigate the germination capacity, field emergence and quality attributes of seed stored under a variety of storage types and used by farmers in the rain fed sorghum producing areas of Sudan as a seed source. Seeds were obtained from a conventional silo with limited controlled conditions, a brick wall warehouse, and a corrugated iron warehouse ( Where seeds were stored for one and two years); a modern silo with controlled conditions and underground pits (Where seeds were stored for one year), and newly harvested seed. The following tests were carried out: 100seed weight, standard germination test, purity test, viability (Tetrazolium) test, oil and protein content, percentage of seeds in different classes of ungerminated seeds, percentage of abnormal seedlings and field emergence. Seeds stored in brick wall warehouse and traditional silo for one year showed significantly lower values of the tested attributes compared to seeds stored in the modern silo and newly harvested seeds. Storing the seeds in these two types of storage for a further year resulted in further significant decreases in the tested characters. Seeds stored in corrugated iron warehouse for one year had significantly lower test results compared to seeds stored in the traditional silo and brick wall warehouse. Storing seeds for a further year in a corrugated iron warehouse resulted in the loss of 60% of their viability and germination capacity. Also, the result revealed that seeds stored in underground pits (matameer) for one year lost most of their germination ability (< 0.11) and their viability (< 0.17) and may contain up to 30% inert matter and other seeds. Seeds stored in the modern silo had protein and oil contents similar to those of the newly harvested seeds, but significantly higher than all other types of storage. The significance of these results was discussed. Keywords: Sorghum, storage, seed quality. INTRODUCTION Sorghum, (Sorghum bicolor (L.) Moench.) is the principal staple food in the Sudan. In Gedarif, the main producing area in the country, depending on rainfall, 2 to 3 million ha of sorghum are grown annually. Productivity is very low not exceeding, on average, 500 kg ha 1. Poor seed quality is perhaps one of the most important factors contributing to this low productivity. In the Sudan, as elsewhere in many African countries, the seed industry is not welldeveloped. Farmers usually keep part of their produce to be used as seed in the following season, this is usually stored in traditional storage facilities which include underground pits (matameer) and different types of warehouses Sometimes farmers may not be able to provide their own seed. In this case a farmer may obtain seeds from the local market or from other farmers. This seed may have been stored in warehouses, silos, or underground pits and may be one to several years old. The normal moisture content of sorghum seed is around 8% to 12% depending on the conditions during seed maturation and storage (Nguyen and Blum, 2004). Seeds of most grain crops, including sorghum, will maintain satisfactory germination and vigour for about one year at moisture content of 12 to 13% under normal warehouse temperatures. When longer storage is needed, seed moisture content should be less than 11% and the temperature should not exceed 20 0 C (Copeland, 2001). Saravanan et al. (2001) reported that the moldinfected seeds of sorghum exhibited highest reduction during storage of one year in 100seed weight, seed size and vigour and there was a considerable decrease in germination after 12 months. Loss of weight and reduction in germination and seedling vigour of

2 germinating seeds due to fungi and insects, especially if moisture content increases during storage, has been reported by some workers (Chitio et al., 2004). In Ethiopia, Dejene et al. (2004) reported that germination of sorghum grain from soil pit decreased by 6% per storage month. The present study investigates germinability and some quality aspects of sorghum seed stored under different storage facilities in the main sorghum producing area of the Sudan. MATERIALS AND METHODS Storage Type The study was conducted during the period Sorghum seed of the local cultivar Feterita were collected from the following storage facilities according to International Rules for Seed Testing (ISTA, 1985): Conventional silo with limited controlled conditions (Temperature C, R.H. 6 14%) used for large scale storage of sorghum, silo with fully controlled conditions (Temperature 20 0C, R.H. 6%), warehouse with corrugated iron walls and roof, fumigated warehouse with brick walls, corrugated iron roof and cement floor, underground pits without lining and newly harvested seed. Storage Duration In warehouses (both brick wall and corrugated iron type) and conventional silo samples were collected from seeds stored for one and two years. In the case of underground pits seeds are usually stored only for one year and, therefore, seed stored for only one year were collected. The controlled conditions silo seeds were available for one year only. Determinations All determinations were carried out according to International Rules for Seed Testing (ISTA, 1985). 100seed weight: 4 x 100 seeds were weighed and expressed in g. Moisture content: 4 x 100 seed samples were ground and ovendried at 130 0C for two hours. Calculations were based on the wet basis. Purity test: 4 x 90 g. samples were sorted into pure seed, other seed and inert matter and the data were expressed as percentages. Germination test: 4 x 100 seeds were germinated in rolled germination towels moistened with water in an incubator at 25 0 C. The first count was made 4 days later and the second 10 days from the start. Types of ungerminated seed: At the end of the germination test, ungerminated seeds were counted and classified into hard, fresh, molded and empty. Tetrazolium test: The test was carried out on samples from the second and third seasons only. The procedure and evaluation were based on the rules issued by AOSA (1978). Oil content: was determined by the Soxhlet extraction method as described by AOAC (1984). Protein content: was determined by the semimicro Kjeldal method as described in Pearson's Chemical Analysis of Foods (Egan et al., 1981). Field emergence: Was studied following the procedure described by Amos (1984) using a completely randomized design. The first seedling count was made after seven days and the second after fourteen days from sowing. Abnormal seedlings were determined according to the description of ISTA (1985). Data analysis Data were statistically analyzed using the analysis of variance (ANOVA) method according to Gomez and Gomez (1984). Means were separated by the Duncan Multiple Range Test (DMRT). Data in the form of percentages were transformed using Arcsin transformation. Correlation and regression analysis was done according to Gomez and Gomez (1984) using computer facilities. RESULTS AND DISCUSSIONS RESULTS Effects of storage type on the measured parameters are given below: Seed moisture content and seed weight Seeds stored in underground pits and those stored in corrugated iron warehouse for one and two years had significantly the highest moisture content than other storage types, but they were not significantly different from each other (table 1). They were followed by seeds stored for two years in the brick wall warehouse. Newly harvested seeds, seeds stored in conventional silo for one and two years and those stored in the controlled conditions silo had significantly the lowest moisture content as compared to other storage types, but were not significantly different form each other.newly harvested seeds, seeds stored for one year in the controlled conditions silo and those stored for one year in the conventional silo recorded a significantly higher weight compared to other storage facilities. Seeds stored in underground pits had significantly the lowest weight compared to seeds from all other storage facilities, followed by seeds stored for one and two years in the corrugated iron warehouse. Seeds stored in a brick wall warehouse for two years maintained a weight similar to that of seeds stored in corrugated iron warehouse for one year. Also, seeds stored in a conventional silo for two years maintained a weight similar to that of seeds stored for one year in brick wall warehouse. Seed purity newly harvested seeds, seeds from silo with controlled conditions and those stored in the 2

3 conventional silo for one year had significantly the highest percentage of pure seed (table 2). Seeds stored for two years in corrugated iron warehouse had the lowest percentage of pure seed. Significant differences were also observed in the percentage of other seeds which contaminated the stored seeds. Other seeds were only present in seeds stored in the brick wall warehouse for two years and in the corrugated iron warehouse for one or two years with the highest percentage found in the latter. The presence of inert matter was observed in all classes of storage with significant differences among these classes. The highest percentage of inert matter was observed in seeds stored in underground pits and those stored in a corrugated iron warehouse for two years. Newly harvested seeds and those in a silo under controlled conditions contained the lowest percentage of inert matter the same as those stored for one year in conventional silo. Table. 1: Moisture content (%) and 100seed weight (g) and germination (%) of seeds stored in Different storage facilities. Storage Type Moisture 100Seed Weight Germination (%) Content (%) (g) 4 DAS 10 DAS Newly harvested 4.0 d 4.0 a 99.0 a 99.5 a Brickwall warehouse (one year) 4.5 c 3.6 b 92.8 b 93.7 b Brickwall warehouse (2years) 4.6 b 3.1 c 80.1 d 81.0 d Corrugated iron warehouse (one year) 4.9 a 4.0 a 63.8 e 64.0 e Corrugated iron warehouse (2years) 4.9 a 2.7 d 3 f 34.0 f Conventional silo (one year) 4.0 d 3.9 a 93.5 b 93.7 b Conventional silo (2years) 4.0 d 3.8 b 87.5 c 88.0 c Controlled conditions silo (one year) 4.0 d 3.9 a 98.5 a 99.0 a Under ground pit 4.9 a c 1 g 1 g Mean Cv % S.E. ± Table 2: Purity test (%) for seeds from the different storage facilities. Types of Storage Pure Seed Other Seed Inert Matter Newly harvested 98.6 a d 1.4 e Brick wall warehouse (one year) 92.7 c d 7.3 c Brick wall warehouse (2years) 7 d 1.0 c 29.0 a Corrugated iron warehouse (one year) 7 d 5.0 b 25.0 b Corrugated iron warehouse (2years) 6 e 1 a 3 a Conventional silo (one year) 98.8 a d 0.2 e Conventional silo (2years) 96.0 b d 4.0 d Controlled conditions silo (one year) 99.9 a d 0.1 e Under ground pit 7 d d 3 a Mean Cv% S.E. ±

4 Table 3: Percentage of seeds in the different classes of ungerminated seeds from the germination test. Types of Storage Hard Seed Fresh Seed Molded Seed Empty Seed Mean Newly harvested f Brick wall warehouse (one year) e Brick wall warehouse (2years) d Corrugated iron warehouse (one year) c Corrugated iron warehouse (2years) b Conventional silo (one year) e Conventional silo (2years) 0 d Controlled conditions silo (one year) f Under ground pit a Mean Cv% 9.81% S.E. ± Germination Significant differences were observed between the germination percentages taken four and ten days under all storage conditions (table 1). Newly harvested seeds and seeds stored in the controlled conditions silo for one year had the highest germination percentage. Seeds stored in underground pits had the lowest percentage of germination. After one year of storage these seeds have lost almost all their germination capacity. Seeds stored in corrugated iron warehouse for one year were distinctly worse compared to other aboveground storage facilities; when stored for a further year they lost more than 50% of their already low germination capacity. Storing seeds for two years in the brick wall warehouse and the conventional silo had significantly reduced germination capacity compared to storing them for one year only. Seeds stored for one year in either a brick wall warehouse or a conventional silo had the same germination capacity. However, storing seeds for a further year in the brick wall warehouse significantly reduced their germination capacity compared to storing them for a further year in the conventional silo. Ungerminated seed The highest percentage of ungerminated seeds was recorded for the seeds stored in underground pits, followed by those stored in corrugated iron warehouse for two years (table 3). A small percentage of ungerminated seeds was observed in the newly harvested seeds, seeds stored in controlled conditions silo and those stored in conventional silo for one year. There were no significant differences in the percentages of ungerminated seeds between seeds stored in the conventional silo for two years and seeds stored in brick wall warehouse for two years. Field establishment, seed viability and abnormal seedlings Seeds stored in the silo with controlled conditions and those stored in conventional silo for one year had the same level of field emergence as newly harvested seeds. They had significantly the highest percentage of field emergence compared to other storage facilities. Seed from the brick wall warehouse were significantly better in field emergence than those from the corrugated iron warehouse. The lowest percentage of field emergence was observed for seeds stored in underground pits (table 4). Newly harvested seeds and seeds stored in the modern silo with controlled conditions gave the highest percentage of viable seeds as estimated by 4

5 the Tetrazolium method compared to seeds from other storage facilities (table 4). The lowest seed viability in all aboveground storage facilities was in the corrugated iron warehouse. On the other hand, most seeds stored in the under ground pits seemed to have lost their viability by the end of the storage period. Protein and oil content The highest percentages of crude protein and oil were recorded for newly harvested seeds, seeds stored in a silo under controlled conditions and those stored in underground pits (table 5). The lowest percentages were recorded for seeds stored for two years in corrugated iron warehouse. Correlation studies There was a high positive correlation between germination and viability as estimated by the Tetrazolium method (R 2 = 0.935) and between percentage germination and field emergence (R 2 = 0.985). DISCUSSION As moisture content of the seeds is one of the most important factors determining seed longevity during storage (Qaisrani, 2000), conditions which maintain seeds moisture content close to that of the newly harvested seed provide better storage. The results of this study indicated that these conditions were provided by the different types of silos. It also suggested that as far as moisture content and germinability are concerned, a conventional silo is as good as the more expensive modern silo kept under controlled conditions. In the storage facilities which provide little protection for stored seeds from exchange of moisture with the surrounding air, namely the corrugated iron warehouse and the underground pits seeds recorded the highest moisture content in this study. This is supported by Lemessa et al. (2000) for underground pits. The results also revealed that a brick wall warehouse is a better facility for storing sorghum seed than a corrugated iron warehouse in terms of moisture content and viability. This may, possibly, be related to the better insulation offered by the brick wall warehouse both in terms of climatic factors and seed infection. However, as seeds in the brick wall warehouse were not completely insulated their moisture content continued to rise during the second year of storage. On the other hand, the moisture of seeds stored in the corrugated iron warehouse seemed to have reached equilibrium with that of surrounding air in the first year and hence no increase was observed in the second year of storage which may again reflect the poor insulation provided by this type of storage. Poor storage conditions resulted in significant decreases in seed weight compared to newly harvested seed and modern storage facilities. Loss in seed weight is associated with seed deterioration and results from the degradation of storage compounds and the leaching of inorganic and organic compound (AbdulBaki and Anderson, 1972), or from seed infection with pathogens or insects (Saravanan et al., 2001; Chitio et al., 2004 ). The two most prevalent types of sorghum seeds storage in sorghumgrowing areas of Sudan are the corrugated iron warehouses and underground pits. Many farmers rely on those sources for their seed supply. Small farmers rely particularly on seeds from underground pits. The result of this study revealed that seed from these two sources had the lowest germination capacity, viability and field emergence compared to other storage facilities and may have, thus, contributed to the poor stands and low yield characteristic of the rainfed sorghum producing areas of Sudan and perhaps other parts of Africa. Seed from the underground pits, in particular, seemed to have lost most of their germination capacity. This result is supported by Lemessa et al. (2000) and Dejene et al. (2004). Storing the seeds in a corrugated iron warehouse for one year drastically reduced their viability and germination capacity so they were not suitable for planting. Storing these seeds for a further year reduced their germination capacity by 50%. Although relative humidity was not measured in this study, it would appear that it played an important in role in the decreased germination of seeds from the corrugated iron warehouses where relative humidity was likely to be high. This view is supported by the report of Qaisrani (2000) and Desai (2004). As for underground pits, it may seem likely that oxygen depletion suggested by Krishnamurthy et al. (1990) may have led to embryo death and hence loss of viability and germinability and poor seedling emergence and establishment. Seeds from the modern storage facilities are subjected to purification such as sieving and blowing and hence their very low percentage of other seed and inert matter. On the other hand, seeds from traditional storage facilities contain a large proportional of inert matter and other seeds sometimes reaching as high as 30%. This would mean that sometimes 30% of the material planted by farmers is not really sorghum seed which may represent a financial loss in terms of the material planted and also the consequent loss in plant population. Storing the seed under controlled conditions in the modern silo maintained seed 5

6 viability, germinability and field emergence at the same level as newly harvest seed. However, in the conventional silo where humidity and temperature were not strictly controlled, seed viability was significantly reduced compared to storing them in a modern silo with controlled conditions. The results of this study also revealed that seed stored for two years in conventional silo were as good as seeds stored in brick wall warehouse for one year in term of seed viability, germinability and field emergence. The results indicated that molded seeds and fresh ungerminated seeds are most prevalent under poor storage conditions. In the corrugated iron warehouse extending the storage period for two years subjected the seeds to a large increase in molds and insects attacks as judged by the large percentage of empty seeds. Insect larvae are reported to feed internally and damage the kernel (Chitio et al., 2004). Desai (2004) stated that sorghum seed is very vulnerable to storage pests and molds and must not be stored in an environment where the moisture content is above 1011%. Protein content is considered as a main biochemical parameter of sorghum grain (Krishnamurthy et al., 1990). The fall in the protein content of seeds from the traditional storage facilities and conventional silo compared to newly harvested seeds, is supported by the report of Reddy and Bushama (1986) who ascribed the decrease of protein quality to insect damage. The decrease in protein content is a manifestation of seed deterioration which is associated with the degradation of storage compounds (AbdulBaki and Anderson, 1972). Bradbeer (1988) considered storage proteins in sorghum seed as an important reserve substance. Oil content was also adversely affected by the poor storage facilities compared to the modern facilities. Furthermore, prolonging the storage period under these conditions led to further decrease in the fat content. The maintenance of fat level in the underground pit probably reflects the early death of embryos. Increase in the percentage of abnormal seeding is one of the physiological manifestations of seed deterioration and may also result from mechanical damage or infection by microorganisms during storage (Perry, 1973). As such, storage conditions, which encourage seed deterioration, are expected to show a higher percentage of abnormal seedlings in the germination test. This was clearly indicated by the results of this study where significantly higher percentage of abnormal seedlings was recorded for the warehouse storage facilities compared to silo storage; with longer storage and, hence, further deterioration, the % of abnormal seedlings significantly increased. The high positive correlation between germination and viability as estimated by the Tetrazolium test may suggest using the test as a reliable indicator of sorghum viability and hence germination capacity. Being a fast test, it would save a lot of time and effort in screening especially large numbers of sorghum samples (Moore, 1973). Also the high positive correlation between the laboratory germination test and field emergence suggests that the test is a good indicator of seed performance under field conditions as was indicated by other workers (Moore 1973; Yaklich and Kulik, 1979). CONCLUSION Storing sorghum seed in silos with either limited or fully controlled conditions maintained seed moisture, viability, germinability and field emergence close to that of newly harvested seed. A brick wall warehouse is a better facility for storing sorghum seed than a corrugated iron one. The two most prevalent types of sorghum seeds storage in rainfed sorghum producing areas of Sudan namely underground pits and corrugated iron warehouses recorded the lowest germination percentage, viability and field emergence. Seeds form underground pits lost almost all their germination capacity while seeds form the corrugated iron warehouse lost more than 60% of their germination capacity after two years of storage. Seeds from traditional storage facilities contain a large proportion of inert matter and other seeds, sometime as reaching as high as 30%. This would mean that sometimes 30% of the material planted by farmers is not sorghum seed with the consequent financial loss in terms of the material planted and the plant population established. 6

7 Table 4 Field emergence (%), viability (Tetrazolium) test (%) and abnormal seedlings of seeds from The different storage facilities. Types of Storage Field Emergence Viable 4DAS 10DAS Seed Newly harvested 98.0 a 99.0 a 99.8 a Abnormal Seedlings 0.7 e Brick wall warehouse (one year) 84.7 c 88.3 c 85.3 c 5.0 c Brick wall warehouse (2years) Corrugated iron warehouse (one year) Corrugated iron warehouse (2years) Conventional silo (one year) Conventional silo (2years) Controlled conditions silo (one year) 8 d 67.3 e 43.5 f 97.7 a 90.7 b 98.7 a 81.0 d 68.0 e 44.0 f 98.0 a 91.3 b 99.0 a 7 d 6 e 5 f 90.8 b 8 c 98.0 a 9.0 b 8.0 b 12.0 a 0.8 e 1.0 e 0.7 e Under ground pit 24.5 g 24.5 g 16.5 g 1.1 e Mean Cv% S.E. ± DAS = Days After Sowing Table 5 Crude protein (%) and oil content (%) of seeds stored in the different storage Facilities. Types of Storage % Crude Protein % Oil Content Newly harvested 16.0 a 3.9 a Brick wall warehouse (one year) 14.6 b 3.6 b Brick wall warehouse (2years) 14.1 b 3.3 c Corrugated iron warehouse (one year) 14.4 b 3.4 c Corrugated iron warehouse (2years) 13.6 c 2.2 d Conventional silo (one year) 14.8 b 3.6 b Conventional silo (2years) 14.4 b 3.5 b Controlled conditions silo (one year) 15.8 a 3.9 a Under ground pit 15.8 a 4.0 a Mean Cv% S.E. ± REFERENCES AbdulBaki, A.A. and Anderson, J.D (1972). Physiological Handbook of Seed Testing, Association of Official Seed Analysts, USA and biochemical deterioration of seeds. In: Kozlowski T.T. (ed.) Seed Biology, vol. 2, Academic Press, New Bradbeer, J.W (1988). Seed Dormancy and Germination. York. pp North Holland Publishing Company, Amsterdam Amos, O.O (1984). Effect of Seed Vigour Levels on Field Performance and Yield of grain Sorghum (Sorghum bicolor (L.) Moench.). Ph.D thesis. Miss. State Univ., Miss. State, MS Association of Official Agricultural Chemists (1984). Official Methods of Analysis of the Association of Official Agricultural Chemists. 14 th Ed. Washington, D.C Association of Official Seed Analysts (1978). Tetrazolium Testing Handbook. Contribution No. 29 to the Chitio, F.M, Pendelton, B.B. and Michels Jr., G.J (2004). Resistance of stored sorghum grain to maize weevil. Int. Sorghum and Millets Newsletter 45: 3536 Copeland, L.O. (2001). Principles of Seed Science and Technology. SpringerVerlag, New York Dejene, M.; Yuen, J. and Sigvald, R (2004). The impact of storage methods on storage environment and sorghum grain quality. Seed Sci. & Technol. 32:

8 Desai, B.B (2004). Seeds Handbook: Biology, Production, Processing and Storage. CRC Press, Boca Raton, Florida, USA Egan, H.; Kirk, R.S. and Sawyer, R (1981). Pearson ' s Chemical Analysis of Foods. 8 th Edn., Longman Scientific and Technical, Harlow, U.K Gomez, K.A. and Gomez, A.A (1984). Statistical Procedures. 2 nd Ed., John Wiley and Sons, New York International Seed Testing Association (1985). International rules for seed testing. Seed Sci. & Technol. 13: 1513 Krishnamurthy, K.; Madani, H.H. and Mohamed, M.E. (1990). Seed Storage and Pest Management. FAO Project Publication, Department of Plant Protection, Ministry of Agriculture, Wad Madani, Sudan Lemessa, F.; Bultosa, G. and Wakari, W (2000). Quality of grain sorghum (Sorghum bicolor (L.) Moench.) stored in traditional underground pits; Case study in two agroclimatic zones in Haraghe, Ethiopia. J. Food Sci. & Technol., Mysore 37: Moore, R.P (1973). Tetrazolium staining for assessing seed quality. In: Heydecher W. (ed.) Seed Ecology, Butterworths, London. pp Nguyen, H.T. and Blum, A (2004). Physiology and Biotechnology Integration for Plant Breeding. CRC Press, Boca Raton, Florida, USA Perry, D.A (1973). Interacting effects of seed vigour and environment on seedling establishment. In: Heydecher W. (ed.) Seed Ecology, Butterworths, London. pp Qaisrani, R. (2000). Safe Storage of Cereals at Higher Moisture Levels. Research report, CSIRO, Stored Grain Research Laboratory, Kangaroo Island, SA Reddy, M.U. and Pushama, P (1896). Effect of storage and insect infestation on thiamine and niacin content in different varieties of rice, sorghum and legumes. Nutrition Reports International 34: Saravanan, T.; Valluvaparidasan, V. and Ravichandran, V (2001). Effect of mold causing organisms of sorghum on seed and seedling vigour. Research on Crops 2: Yaklich, R.W. and Kulik, M.M (1979). Evaluation of vigour tests in soybean seeds: Relationship of the standard germination test, seedling vigour classification, seedling length and Tetrazolium staining to field performance. Crop Sci. 19:

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