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SEED PRIMING IMPROVES EARLY SEEDLINGGROWTH AND NUTRIENT UPTAKE INMUNGBEANHamidullah Shah a , Tabinda Jalwat a , Muhammad Arif b & GhazalMiraj aa Department of Agricultural Chemistry , NWFP AgriculturalUniversity , Peshawar , Pakistanb Department of Agronomy , NWFP Agricultural University ,Peshawar , PakistanPublished online: 03 Apr 2012.

To cite this article: Hamidullah Shah , Tabinda Jalwat , Muhammad Arif & Ghazal Miraj (2012) SEEDPRIMING IMPROVES EARLY SEEDLING GROWTH AND NUTRIENT UPTAKE IN MUNGBEAN, Journal of PlantNutrition, 35:6, 805-816, DOI: 10.1080/01904167.2012.663436

To link to this article: http://dx.doi.org/10.1080/01904167.2012.663436

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Journal of Plant Nutrition, 35:805–816, 2012Copyright C© Taylor & Francis Group, LLCISSN: 0190-4167 print / 1532-4087 onlineDOI: 10.1080/01904167.2012.663436

SEED PRIMING IMPROVES EARLY SEEDLING GROWTH

AND NUTRIENT UPTAKE IN MUNGBEAN

Hamidullah Shah,1 Tabinda Jalwat,1 Muhammad Arif,2 and Ghazal Miraj1

1Department of Agricultural Chemistry, NWFP Agricultural University, Peshawar, Pakistan2Department of Agronomy, NWFP Agricultural University, Peshawar, Pakistan

� A series of greenhouse and field experiments were conducted at Northwest Frontier Province(NWFP) Agricultural University, Peshawar, Pakistan to study the effect of seed priming with dilutesolutions of phosphorus (P) on seedling growth and nutrient uptake of mungbean (Vigna radiataL.). Seed of mungbean cv. ‘NM-92’ were primed in 0.01% and 0.02% P solutions and also inwater for four hours along with a non-primed seed (control). The primed seed was washed withdistilled water to remove nutrients adhering to seed coat and air dried. The amount of nutrientsintrogressed into seed or adhered onto the seed hull was measured. Higher P was introgressed by theseed primed with 0.02% P solution followed by seed primed in 0.01% P solution as compared towater soaked and dry seeds both in nonwashed and surface washed seeds. Likewise, seed primingwith P solutions significantly improved seedling P concentration and uptake 14 and 21 days aftersowing as compared to water soaked and dry seeds in both greenhouse and field experiments. Simi-larly, seed priming with P solutions significantly improved fresh and dry weight and plant heightof mungbean seedlings 21 days after sowing in the field experiment.

Keywords: seed priming, phosphorus, uptake, seedling growth

INTRODUCTION

Mungbean (Vigna radiata L.) or green gram is a tropical legume and itsseeds are primarily used for food purposes; it is a rich source of lysine andprotein, and thus can supplement a cereal-based human diet. The whole orsplit seed is usually cooked as dhal or boiled with rice (Rosaiah et al., 1993,Singh and Singh, 1992). The seed may be milled and ground into flour formaking noodles, breads, and soups. The spice-roasted seeds of mungbeanare sometimes eaten as a snack food (Malik, 1994). In the rural areas theimmature green pods are also used as a vegetable. Mungbean stalks, leavesand husk constitute a significant proportion of livestock feed. After picking

Received 5 January 2010; accepted 13 June 2011.Address correspondence to Muhammad Arif, Department of Agronomy, NWFP Agricultural

University, 25000 Peshawar, Pakistan. E-mail: marifkhan75@yahoo.com

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the pods, the whole plant may be ploughed into the soil to improve itsfertility.

In Pakistan, mungbean was grown on an area of 219.2 thousand hectareswith a total annual production of 104.5 thousand tones both under rainfedand irrigated conditions, whereas in Northwest Frontier Province (NWFP)area under mungbean cultivation is 9.5 thousand hectares with a correspond-ing production of 6.1 thousand tones (MINFAL, 2008). It is mostly grownon rainfed lands and thus yields are generally low and variable due to sparse,erratic rainfall and marginal soils.

Phosphorus (P) deficits have an effect on more than 40% of tropicalsoils (Smithson and Giller, 2002) and P is the second most limiting nutri-ent, after nitrogen (N), for crop production. Although N is also necessaryin large amounts for agricultural crops, this N can partly be supplied in asustainable manner through N2-fixing crops (Bationo and Ntare, 2000). Incontrast, P deficiency can only be rectified by bringing in fertilizers fromoutside or by enhancing P mobilization and plant acquisition from the soiland this therefore represents a more serious long-term concern (Sinclairand Vadez, 2002). As the supply of P fertilizers in most developing countriesis limited by a range of constraints, alternative plant-based strategies are re-quired to optimize which fertilizers are available alongside intrinsic soil Preserves (Gandah et al., 2003; Schlecht et al., 2006). Therefore, althoughP may be present in soil, most of it remains in forms relatively unavailablefor plant uptake, and plants consequently suffer acute P deficiency and pro-duce poor yields. Low fertilizer-use-efficiency and losses to the environmentare major worldwide environmental problems and research must seek toimprove fundamental efficiencies of crop nutrient use (Tilman et al., 2002;Sanchez, 2002).

Crop production is widely limited by poor stand establishment and nu-trient deficiencies (Jones and Wahbi, 1992). Particularly in drought proneenvironment, the emergence tends to be irregular and can extend over longperiods (Bougne et al., 2000). The resulting poor stand leaves gaps in thecanopy, which are filled by weeds that compete with the crop plants for thelimiting nutrients (Kropff and Van Laar, 1993). Poor crop establishment isoften cited as a major constraint for mungbean production and high yieldshave been correlated positively with early seedling vigor (Naseem et al.,1997). Healthy plants with well-developed root system can more effectivelymobilize limiting nutrients from the soil and can better withstand adverseconditions. A vigorous early seedling growth has been shown to be associatedwith higher yields (Harris and Jones, 1997).

Crop production is affected by low chemical availability of P and zinc (Zn)(Ryan, 1997). Especially during the early growth stages, a lack of P retardsseedling growth, rendering the young plantlets particularly sensitive to thefrequently encountered dry spells (Jones and Wahbi, 1992). A rapid estab-lishment of healthy seedling and a sufficient supply with P are prerogative

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to reduce the risk of crop failure (Brown et al., 1987). Nutrient priminghas been proposed as a novel technique that combines the positive effects ofseed priming with an improved nutrients supply (Arif et al., 2005; Al Mudarisand Jutzi, 1999). In nutrient priming, seeds are pretreated in solution con-taining the limiting nutrients instead of being soaked simply in water.

This research was therefore conducted to develop a nutrient primingapproach to foster mungbean seeds for the marginal growing conditions ofpoor stand establishment and P deficiencies and enhance yield.

MATERIALS AND METHODS

To study the effects of seed priming on nutrient uptake and seedlinggrowth of mungbean, greenhouse and field experiments were conductedat Northwest Frontier Province (NWFP) Agricultural University, Peshawarduring 2005.

Experimental Site

Peshawar has a warm to hot, semi-arid, sub-tropical, continental climatewith mean annual rainfall of about 360 mm. Summer (May–September) hasa mean maximum temperature of 40◦C and mean minimum temperatureof 25◦C. Winter (December to the end of March) has mean minimum tem-perature of 4◦C and a maximum of 18.4◦C. The average winter rainfall ishigher than that of summer. The highest winter rainfall has been recordedin March, while the highest summer rainfall is in August. Soil of the experi-mental site is deficient in N, P, and available Zn, but has adequate potassium(K). Canal water is available for irrigation (Harris et al., 2007). The physio-chemical properties of the experimental site are given in Table 1.

TABLE 1 Physico-chemical properties of the experimental site

Soil property Experimental site values

Sand (%) 8.7Silt (%) 51.3Clay (%) 40.0Textural class Silty clay loamOrganic matter (g kg−1) 0.845Total N (g kg−1) 0.04CaCo3 (%) 14.4pH 1:1 water 8.02Electrical conductivity (dS m−1) 0.87AB-DTPA extractible nutrientsP (mg kg−1) 3.80K (mg kg−1) 105Zn (mg kg−1) 0.86

Data from Bhatti (2002) and Tariq et al. (2002) for Agricultural University Peshawarfarm.

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D A

C B

FIGURE 1 Sample distribution of the tub for the treatments.

Seed Priming

Seed of mungbean was primed with 0.01% and 0.02% P solutions andalso in water for four hours. Non-primed seed (dry seed) was also included asa control treatment. Potassium dihydrogen phosphate (KH2PO4) was usedas source of P. The primed seed was washed with distilled water to removenutrients adhering to seed coat. After priming, the seeds were air-driedfor 15 minutes to ensure clump free placement/sowing in further experi-ments. The amount of nutrients introgressed into seed or adhered onto theseed hull were quantified by subtracting the nutrient content washed fromthose of non-washed seed. The differences between non-primed and washedprimed seed represented the amount of nutrient introgressed into the seed.

Greenhouse Experiment

The seeds were primed with 0.01% P, 0.02% P solutions, and waterfor four hours. Dry seed was included as a control treatment. Fifty kg ofprepared soil was taken in four tubs and were irrigated prior to sowing. Eachtub represented a replication. At filed capacity, surface soil was hoed andarea was divided into four equal portions (A, B, C, D); each part for sowing12 seeds per treatment (Figure 1 and Table 2). After sowing, all the seedwere irrigated after emergence. Seedlings from all the tubs were harvested14 and 21 days after sowing (DAS).

TABLE 2 Layout of the greenhouse experiment

Phosphorus (P)

R1 R2 R3 R4

A = water A = 0.01% P A = 0.02% P A = non-primedB = 0.01% P B = 0.02% P B = non-primed B = 0.01% PC = non-primed C = water C = 0.01% P C = waterD = 0.02% P D = non-primed D = water D = 0.02% P

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Field Experiment

For this experiment, the field was irrigated and at proper moisture con-dition, it was thoroughly ploughed using cultivator followed by planking.Mungbean variety ‘NM-92’ was sown at a seed rate of 25 kg ha−1 in a plotsize of 5 m by 5 m consisting of ten rows five meter long and 50 cm apart.Nitrogen was applied at the rate of 20 kg ha−1 as a starter dose using urea assource. Phosphorus was applied at the rate of 60 kg ha−1 using single superphosphate (SSP) as a source. No irrigation water was applied till 21 DAS.Seedlings from these plots were also harvested 14 and 21 DAS.

Fresh Shoot Weight (g Plant−1)

Ten seedlings from each treatment were harvested 14 and 21 DAS, andweighed for fresh shoot weight. The data were then converted to fresh shootweight per plant.

Shoot Height (cm)

The shoot height of ten randomly selected plants was measured fromthe ground level to the tip of leaf and then averaged.

Dry Weight (g Plant−1)

Data on dry shoot weight were recorded by keeping the fresh samples inoven at 70◦C for 24 h in paper bags and weighed and then converted to dryweight per plant.

Nutrient Uptake (mg kg−1)

Nutrient uptake for the seedling was calculated by using the followingformula:

Nutrient uptake = total dry matter × nutrient concentration

Nutrient (P) Concentration (mg kg−1)

All the dried samples of shoot were grinded and dried. The powderedsamples were then first digested with nitric acid and perchloric acid andthen the aliquots were used for the determination of phosphorus by spec-trophotometer (A.O.A.C., 1990).

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Statistical Analysis

The data were statistically analyzed by analysis of variance appropriatefor randomized complete block design. Means were compared using LSDtest at 0.05 level of probability (Steel and Torrie, 1980).

RESULTS AND DISCUSSION

Phosphorus (P) concentration in seeds increased with increase in Pconcentration of priming solutions. Higher P was introgressed by the seedprimed with 0.02% P solution followed by seed primed with 0.01% P so-lution as compared to dry seed and water soaked seed both in non-washedand surface washed seeds. Phosphorus concentration decreased with surfacewashing at 0.395 and 0.337 mg g−1 in seed primed with 0.01 and 0.02% P so-lutions, respectively, which indicated that these amounts of P were adheredto the seeds coat of the seed (Table 3).

Likewise, seed priming significantly improved P concentration and up-take of the mungbean seedlings. Higher P concentration was measured inthe seedling obtained from the plot sown with seed primed with 0.02% Psolution followed by 0.01% P solution 14 and 21 days after sowing (DAS)as compared to dry seed and water soaked seed in both tub and field ex-periments. Plant uptake was not significant both 14 and 21 DAS in tubexperiment but was significant in the field experiment. Seed priming in Psolutions significantly improved P uptake both 14 and 21 DAS in the fieldexperiment. Phosphorus uptake was significantly higher in seed primedwith 0.01 and 0.02% P solutions than dry seed and water soaked seed(Figures 2 and 3). This shows that uptake of nutrients was determined bythe rate of nutrient supplied to the seed surface. It is also worth mentioningthat water soaked seed performed better than dry seed in field experimentwhich indicates the beneficial effects of water soaking.

In similar fashion, seed priming significantly improved fresh and dryweight and plant height of mungbean seedlings 21 DAS in the field ex-periment. Higher fresh and dry weights were recorded in plots sown with

TABLE 3 Effect of seed priming on P concentration (mg g−1) of washed and non-washed seeds

P concentration P concentrationTreatment of imbibed seed of surface washed seed Difference

Non-primed (control) 3.300 c 3.305 c 0.005P 0.01% 7.250 b 6.855 b 0.395P 0.02% 7.480 a 7.143 a 0.337

LSD 0.1678 0.0100

Means of the same category followed by different letters are significantly different from one anotherat P = 0.05 using LSD test.

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Seed priming treatments

NP Water 0.01 % P 0.02 % P

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FIGURE 2 Effect of seed priming with phosphorus (P) solutions on seedling P concentration 14 and21 days after sowing in A) greenhouse and B) field experiments. Vertical bars denote SE.

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FIGURE 3 Effect of seed priming with phosphorus (P) solutions on seedling P uptake 14 and 21 daysafter sowing in A) greenhouse and B) field experiments. Vertical bars denote SE.

seed primed in 0.01 and 0.02% P solutions than dry seed and water soakedseed but water soaked seed performed better than dry seed. Seed primedin 0.01 and 0.02% P solutions resulted in taller plants than dry seed andwater soaked seed which were statistically on par with each other. However,

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TABLE 4 Effect of seed priming with P solutions on fresh and dry shoot weight (g plant−1) ofmungbean seedlings 14 and 21 DAS in greenhouse experiment

Fresh Fresh Dry Dry Plant Plantshoot weight shoot weight shoot weight shoot weight height height

Treatment (14 DAS) (21 DAS) (14 DAS) (21 DAS) (14 DAS) (21 DAS)

NP 0.73 a 1.41 a 0.24 a 0.23 a 16.12 a 17.81 aH2O 0.63 a 1.40 a 0.24 a 0.23 a 16.12 a 18.58 a0.01% P 0.76 a 1.30 a 0.26 a 0.26 a 16.02 a 17.35 a0.02% P 0.70 a 1.58 a 0.26 a 0.26 a 16.35 a 19.50 a

LSD ns ns ns ns ns ns

ns = non-significant, NP = non-primed (control).

seed priming did not significantly improve fresh and dry weight of mung-bean seedlings both 14 and 21 DAS in the greenhouse experiment (Tables 4and 5).

There are several advantages of using seed priming to deliver micronu-trients to seeds: the effects of uneven application of zinc to the soil areavoided as each seed is exposed to the nutrient; some uptake is guaranteed;the amounts required are likely to be orders of magnitude less than for soilapplication (Harris et al., 2008). These results indicated that P added toseed by priming are not removable by surface rinsing and hence like to havebeen introgressed into the seed. These results are in line with the findingsof Ajouri et al. (2004) who reported that priming seed of barley with P andZn solutions increased the content of P and Zn in the seeds and on theseed coats. Similar results were observed by Asgedom and Becker (2001).In our results, the application of low rate of nutrient to seeds increasedtheir concentration and could be effective in reduction of fertilizer doseand cost. The results are supported by the findings of Lorenz (1994) whoreported that zinc application to seed enhanced zinc concentration of maizetissue significantly above control. Similarly, Savethri et al. (1999) and Singh

TABLE 5 Effect of P and water priming on fresh shoot weight (g plant−1) of mungbean seedlings 14and 21 DAS in field experiment

Fresh Fresh Dry Dry Plant Plantshoot weight shoot weight shoot weight shoot weight height height

Treatment (14 DAS) (21 DAS) (14 DAS) (21 DAS) (14 DAS) (21 DAS)

NP 1.00 a 3.023 b 0.14 a 0.393 c 19.27 a 27.06 bH2O 1.10 a 3.382 b 0.16 a 0.418 bc 20.42 a 26.43 b0.01% P 1.09 a 4.188 a 0.13 a 0.519 ab 19.96 a 30.29 a0.02% P 1.13 a 4.653 a 0.16 a 0.577 a 19.95 a 30.70 a

LSD ns 0.6749 ns 0.112 ns 2.904

Means of the same category followed by different letters are significantly different from one anotherat P = 0.05 using LSD test. ns = non-significant, NP = non-primed (control).

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and Singh (1973) have used seed soaking as a treatment to increase the Pconcentration of seedling significantly.

Seed priming has been shown to improve early seedling growth andstand establishment (Arif et al., 2005). The improved seedling fresh and dryweight of mungbean may be due to optimum availability of P to seedlingimmediately after their emergence that resulted in enhanced fresh and dryweight. The results are in line with Arif et al. (2005) who reported that seedprimed with 1% P + 2% Zn solution produced higher seedling fresh anddry weight. They also concluded that nutrient (P and Zn) and water primingproduced superior seedling fresh and dry weight in maize. The results agreewith Zhang et al. (1998) who reported that soaking seed in P solution beforeseeding improved the P nutrition of rice during early growth stage. Earlierstudies on rice also suggested that seed coating with Zn or the dipping of theroots of seedling in Zn solution is more effective than surface application ofZn (Giordano and Mortvedt, 1973).

Likewise, several authors have described positive responses to seed prim-ing with water alone in Pakistan and elsewhere (Harris et al., 1999, 2002,2004; Rashid et al., 2002) and the data from this study confirm that sim-ple seed priming with water is an effective way to improve seedling growthand nutrient concentration and uptake in mungbean. This is important forresource-poor farmers as it is a low- or no-cost technology and requires fewexternal inputs.

CONCLUSION

It is concluded that seed priming with 0.01 and 0.02% P solution im-proved fresh and dry weights, plant height and P concentration and nutrientuptake of mungbean seedling. Seed soaking simply in water also performedbetter than dry seed.

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