Research Article

Influence of Seed Priming on Germination and Seedling Vigour of Wood Apple (Feronia limonia Swingle)

Sayan Sau, Binayak Pal, Sukamal Sarkar and Tanmoy Sarkar

  • Page No:  128 - 136
  • Published online: 18 Apr 2019
  • DOI : HTTPS://DOI.ORG/10.23910/IJBSM/2019.10.2.1967

  • Abstract

The present experiment was conducted to study the influence of seed priming treatments on germination and seedling vigour of wood apple (Feronia limonia Swingle) at Horticulture experimental and learning unit, Faculty of Horticulture, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia, West Bengal, India (22°5643''N, 88°32'03 ''E and 397 m above mean sea level) during December to June of 2016-2017. The treated seeds were raised on polythene bags under shade net house (covered with 200-micron thickness polythene shade net) condition to avoid climatic irregularities. The experiment was laid out in a completely randomized design comprising eight treatments (GA3 50 ppm and 100 ppm, KNO3 50 ppm and 100 ppm, Thiourea 50 ppm and 100 ppm, water soaking, and directly sown seed i.e. control) and replicated for five times. The result of the present investigation revealed that amongst the different concentrations of used chemicals, GA3 100 ppm was most efficient to ensure the maximum germination percentage (81.67%) which was 96.01% higher than the control. Similar trend of obtaining highest values from GA3 treated seed on most of the seedling growth parameters was also observed such as highest seedling height (442.23 mm), shoot (2.51 mm) and root diameter (3.22 mm), leaf number (12.00), seedling fresh (2.14 g) and dry weight (1.01 g), leaf chlorophyll content [chlorophyll a (78.51 mg g-1 FW), chlorophyll b (98.70 mg g-1 FW) and total chlorophyll (135.70 mg g-1 FW), respectively] and leaf nitrogen (3.40%) and potassium content (3.42%).

Keywords :   Wood apple, germination, seed priming, seedling vigour

  • Introduction

    Wood apple (Feronia limonia Swingle), the only species of genus Feronia (family Rutaceae), is one of the most medicinally rich underutilized fruit. Though, India and Sri Lanka is considered as native place of wood apple, well-managed orchard is rare in both the countries but road side and field-edge planting is common. With the passage of time, cultivation of wood apple spread over Southeast Asia, including northern Malaysia and Penang Island (Hossain et al., 1994; Islam et al., 2006). Dual purpose use of wood apple i.e. raw as well as processed products makes it more economically preferred commodity. Pulp of wood apple is loaded with immense quantity of medicinally important compounds like umbelliferol, dictamnine, xanthotoxol, sco-parone etc. those could be used in the pharmaceuticals industries. From ancient time, in India the ripe fruits are used as a liver and cardiac tonic, while unripe ones as an astringent for checking diarrhoea and dysentery. In addition, the fruits serve as effectual treatment for sore throat, hiccough, and diseases of the gums (O’Neill and Lewis, 1993). The fruit pulp is also used for the preparation of chutney, fruit bar, jam, jelly, and ready-to-serve beverages (Vidya and Narain, 2011).

    Seed propagation in fruit plants are well acknowledged where seed is the only means of multiplication. Though, commercial method of propagation in wood apple is chiefly through seeds, vegetative propagation by budding, cutting, softwood grafting (Raghavendra et al., 2011), and inarching is also successful. Nevertheless, decline in seed viability is noticed within a very short period following its (seed) extraction from fresh fruits of wood apple. As a consequence, low seed viability along with poor germination is encountered as a severe menace during commercial cultivation of wood apple. Such low viability coupled with poor germination occurs presumably due to the physiological event of low metabolic activity referred as ‘dormancy’ or ‘quiescence’ that is associated with hard seed coat, immature embryo or with the result of endogenous chemical germination inhibitors (Wareing and Phillips, 1981; Sharma et al., 2014).

    Multiple researches have been conducted on pre-germination treatments of seeds with different seed priming chemicals (GA3, KNO3, thiourea, salicylic acid, hot water as well as cow dung slurry) in different fruit crops like custard apple, passion fruit, fig, yoshino cherry, papaya etc (Farooq et al., 2005; Basra et al., 2007; Kim, 2019). These chemicals and multiple other seed priming treatments improve seed germination and seedling growth mainly by reducing internal growth inhibitor content (Agrawal and Dadlani, 1995; Hassini et al., 2017) and increasing the growth promoter concentration in seed; improving entry of water into the cell (Arteca, 1996; Sharma et al., 2014) and improving enzymatic activity to initiate several metabolic processes (Maiti et al., 2011).

    In breeding programmes, wood apple seeds are the key component for developing hybrid plants to enrich genetic diversity, and to produce quality rootstock plant (Morton, 1987). It has been proven that seed priming is one of the competent approaches to obtain uniform and healthy seedling but exhaustive information of this approach in wood apple is still scarce. Keeping these facts in concern, the present experiment was conceived to study the effect of seed priming on germination and seedling vigour of wood apple.

  • Materials and Methods

    2.1.  Experimental site and details of seed priming techniques

    The experiment was carried out at the shade net house (covered with 200-micron thickness polythene shade net) of Horticulture experimental and learning unit, Faculty of Horticulture, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia, West Bengal, India (22°5643''N, 88°32'03 ''E and 397 m above mean sea level) during December to June of 2016-2017 . Fresh healthy seed of wood apple were collected from Horticulture Research Farm, Palli Siksha Bhavana, Visva Bharati (identified and validated by Dr. Prahlad Deb, Department of Crop Improvement, Horticulture and Agricultural Botany, Palli Siksha Bhavana (Institute of Agriculture), Visva Bharati, Sriniketan, West Bengal, India) and stored for 45 days in ploythelene bags, and then these stored seeds were used for this experiment. The experiment was laid out in completely randomized design (CRD) having eight distinct seed priming treatments (GA3 50 and 100 ppm, KNO3 50 and 100 ppm, Thiourea 50 and 100 ppm, water soaking, and one control i.e. directly sown seed), replicated five times. To allow the seed priming treatments, 100 numbers of seeds for each treatment were poured in to a large sized Petridis containing 300 ml of respective priming solution for 24 h except the control. Primed seeds after removal from solution immediately allowed to air drying for half an hour and then sowed in 8 × 10cm size polythene bags(of 200 µM) filled with potting mixture of soil+FYM (farm yard manure)+coir peat+sand (1:1:1:1; w/w). The potting mixture was sterilized with Blitox solution (2 g l-1 of water) before sowing of seeds to prevent the diseases.

    2.2.  Observations recorded

    2.2.1.  Germination percentages

    Germination percentages were calculated as the average of three replicates of 100 seeds upto 30 days from the day of seed sowing. The germination percentage was calculated using the following equation by Maguire (1962):

    Germination percentage=(Number of seeds germinated/Number of seeds sown)×100

    2.2.2.  Growth parameters

    Growth parameters such as shoot and root length; shoot and root diameter, Number and length of root laterals, number of leave were assessed at monthly interval upto 90 days from date of seed sowing. Length and diameter determined with the help of digital slide calipers (6”/150 mm, accuracy §0.02 mm, LR44, 2006/66/EC). Seedling fresh and dry weight measured using digital weighing balance (1 mg to 100 g precision). The percent tissue water content (TWC %) of the seedlings was determined using the following equation by Black and Pritchard (2002) :

    TWC (%)=(Fresh weight-Dry weight)/Fresh weight×100

    2.2.3.  Seedling vigor Index

    Seedling Vigor Index-I (SVI-I) and Seedling Vigor Index -II (SVI-II) were calculated with using the following equations given by Kharb et al. (1994):

    Seedling vigor-I=(Seedling length×Germination %)/100

    Seedling vigor-II=(Seedling dry weight×Germination %)/100

    2.2.4.  Leaf chlorophyll content

    To determine chlorophyll content (chlorophyll a, b and total chlorophyll) fresh leaves of 0.5 g was used and soaked in 10 ml methanol (85%) for 24 h, in the presence of little amount of Na2CO3; after that homogenized and centrifuged. Then the optical density of supernatant was spectrophotometrically measured, and chlorophyll content calculated using following formula as described by Normal (1982):

    Chlorophyll "a" (mg g-1 fresh weight)= {12.7(A663)-2.63(A645)}×{v/(1000×W×a)}

    Chlorophyll "b" (mg g-1 fresh weight)= {22.9(A645)-4.68(A633)}×{v/(1000×W×a)}

    Chlorophyll (mg g-1 fresh weight)= {20.2(A645)-8.02(A633)}×{v/(1000×W×a)}

    Where, A = Absorbance at specific wave length (663 and 645 nm); V=Final volume of the chlorophyll extract (ml); W=Fresh weight of the sample (g); a=Path length of light (1 cm)

    2.2.5.  Leaf mineral composition analysis

    To determine leaf N, P, and K content 50 leaves from each replication collected at seedling age of three month. These leaves were rinsed with tap water, followed by 0.2% (v/v)Teepol® solution, 0.1 N HCl, and finally cleaned with double-distilled water. The surface sterilized leaf samples were individually arranged in labelled paper bags and dried in a hot air oven at 70 °C temperature. Then, the dried samples were ground and digested in wet di-acid using nitric acid (HNO3) and perchloric acid (HClO4). The digested materials were then diluted and filtered through Whatman No.1 filter paper and used to determine N content in leaves with the aid of Digestion Block method (Bremner, 1965), P content by vandomolybdophosphoric yellow colour method (Jackson, 1973), K contents with a microprocessor-based flame photometer (Jackson, 1980).

    2.3.  Statistical analysis

    Results are represented in tables and figures as the means of five replicates. Data were subjected to one-way analysis of variance with the GenStatSoftware ver. 19.1(VSNI, UK) and all means of physical and bio-chemical properties were compared using Duncan’s Multiple Range test. Significant differences were assessed at the p≤0.05 probability level.

  • Results and Discussion

    Application of chemicals for seed priming, significantly (p≤0.05) affects the germination percentage of wood apple seeds in comparison to control (directly sown) (Figure 1).

    The highest number of germination percentage (81.67) was recorded from the seeds that were treated with GA3 100 ppm, followed by Thiourea 50 ppm (76.67), which was 96.01% and 84.01% higher than the control, respectively (Figure 2).

    This improvement in germination of wood apple seeds might be due to the stimulating effect of imbibitions on seed germination caused by increased water absorbing capacity (Cho and Lee, 2018). It might be also because GA3, which is involved in the activation of cytological enzymes, stimulates the α-amylase enzyme that in turn transforms insoluble starch into soluble sugars, and it also sets off the radical growth by eliminating some metabolic blocks (Babu et al., 2010). Another physiological basis on favouring germination with GA3 priming is that it also acts directly on embryo to break the seed dormancy, through the promotion protein synthesis and elongation of coleoptiles and leaves and helps in the production of ethylene which invokes the synthesis of hydrolases, especially amylase (Stewart and Freebarin, 1969). Similar results were obtained by Athani et al. (2013) in guava; Singh et al. (2001) in jujube; Moustafa and Al Zidgali (1995) in citrus and Rajput et al. (1999) in wood apple, jujube and Manilkara sp.

    Data in Table 1 and 2 represent significant variation in shoot,

    root and seedling length of wood apple upon exposure to various seed priming treatments. A gradual progression in both root and shoot elongation was recorded up to the 3rd month, from the date of seed sowing. The maximum length of shoot (7.43 mm), at the 1st month was observed in the seedlings developed from the GA3-(100 ppm)-treated seeds, which was statistically at par with that of the Thiourea-(50 ppm)-treated seeds. However, at the transplanting stage (i.e. 3rd month growth stage of the seedlings),the resultant shoot lengths showed a non-significant variation irrespective of treatments except the control (non-primed/directly sown seed), though the maximum shoot length (118.92 mm) was observed in the seedlings that were raised from GA3 (100 ppm)-primed seeds. In case of root length, in the 1st month the maximum length (157.46 mm) was observed in the seedlings that were developed from seeds treated with Thiourea 100 ppm, which was statistically at par with rest of the other treatments, except GA3 50 ppm and control (non-primed/directly sown seed); and with the progression of time, at 3rd month, the maximum root length (327.06 mm) was recorded in seedlings that received the seed priming treatment i.e. GA3 100 ppm. Similar trends were also recorded in case of total seedling length in different months, and the highest seedling length at the 3rd month stage was recorded from the seedlings that were primed with GA3 100 ppm, which was 85.21% more lengthy than the control ones.

    Effect of seed priming on shoot and root diameter, as shown in Table 3 revealed that gradual increasement occurred from the 1st to 3rd month in both the cases. The seeds, treated with 100 ppm GA3, 50 ppm KNO3, 100 ppm KNO3 or 100 ppm Thiourea-treated exhibited comparable results (based on shoot diameter) with the maximum values (1.52 mm,1.48 mm, 1.44 mm, and 1.46 mm, respectively) recorded at the 1st month of germination. On the other hand, seedlings developed from directly sown seeds recorded minimum (1.09 mm) value (of shoot diameter). At the 3rd month, maximum shoot diameter (of 2.51 mm) was observed in the seedlings that received priming treatment of GA3 100 ppm followed by KNO3 50 and 100 ppm. In case of root diameter, in the 1st month, a non-significant variation was observed among the seedlings that were developed from different priming treatments, whilst at the 3rd month the maximum root diameter of 3.22 mm was recorded in seedlings that were developed from seeds primed with GA3 100 ppm followed by KNO3 100 ppm (3.16 mm).The minimum values were recorded in those seedlings that were developed from directly sown seeds i.e. control.

    Data in Table 4 reveals significant improvement in the number of root laterals and leaves of wood apple seedlings when subjected to different seed priming treatments. In the 1st month, the highest number of root laterals were recorded from priming treatment of Thiourea 100 ppm which was statistically at par with KNO3 50 ppm; and the lowest results were obtained from the directly sown seeds.

    In the 3rd month similar results were recorded as well, where in, the maximum number of root laterals were obtained from GA3 100 ppm (42.67), Thiourea 100 ppm (43.33) and Thiourea 50 ppm (42.33), respectively. During the observation period of 90 days, a result of monthly gain of 2-4 fully expanded leaves was recorded in wood apple. Priming treatment with GA3 100 ppm and KNO3 100 ppm recorded a total number of 12 leaves at the 3rd month stage, which was 38.40% more than that of the seedlings which were developed from directly sown seeds.

    The increase in growth parameters like length, diameter of shoot and root, with treatment with GA3 might be because this plant growth regulator enhanced the osmotic uptake of nutrients, thereby causing greater cell division and elongation (Sen et al., 1990). GA3 treatment is also responsible for production and translocation of photosynthates across phloem tissue to the root zone, which might be accountable for enhancing the root growth. The increase in the number of leaves might be due to the activity of GA3 at the apical meristem, ensuing in more synthesis of nucleoprotein that is responsible for increase in the frequency of leaf initiation (Sen and Ghunti, 1976). The observation analogues to the earlier findings of Kumari et al. (2007) and Manekar et al. (2011) in aonla; Anburani and Shakila (2010) in papaya; Patil et al. (2012) in Rangpur lime; Nimbalkar et al. (2012) in karonda.

    According to data presented in Table 5,

    seed treatment of wood apple showed a significant (p≤0.05) effect on the seedling fresh weight, dry weight and tissue water content of wood apple when compared to control (directly sown). Both the maximum values for fresh and dry weight of seedlings (2.14 g) were obtained from treatment with GA3 100 ppm, followed by KNO3 100 ppm, whilst the minimum values were recorded from the control ones. In accordance with the previous result, though the lowest tissue water content was observed in the same treatment GA3 100 ppm yet, no significant variation among the priming treatments were recorded, with the only exception of those seedlings that were developed from directly sown seeds, which in fact recorded the highest tissue water content (63.37%). It might be due to the overall growth of the seedlings and an increased rate of photosynthesis that led to the overall assimilation and redistribution of photosynthesis within the seedlings and hence, resulted in higher fresh and dry weight, and TWC. Thus, increased growth is a consequence of increased dry matter accumulation. The results are in close conformity with the findings of Pampanna and Sulikeri (1999) in sapota; Meena and Jain (2005), Sasikala and Srimathi (2006) in papaya.

    Figure 3 (A and B) reflects that seed priming treatment had a significant effect on seedling vigour index (both I and II) of wood apple.

    The increase in vigor index has been parallel to the corresponding enhancement of germination percentage for all the seed priming treatments studied here. Highest values (312.12) for vigour index I and (0.70) vigour index II were obtained from GA3 100 ppm primed seeds, whereas, lowest values for both the cases were recorded in the control treatment. The highest seedling vigour in GA3-primed germinated seeds is presumably due to enlargement of embryos, higher rate of metabolic activity and respiration, improved utilization and mobilization of metabolites to growth points and higher activity of enzymes. Enzymatic and hormonal mechanisms stimulate metabolic processes such as sugar mobilization, protein hydrolysis, oxidation etc. (Earplus and Lambeth, 1974), which leads to an increase in root length, shoot length, and seedling dry weight that in turn increases seedling vigour. The present results are in conformity with the results of Gurung et al. (2014).

    Priming of wood apple seeds significantly influences the leaf chlorophyll contents (Figure 4) as well as the mineral composition of leaf (Table 6).

    The maximum values of chlorophyll a (78.51 mg g-1 FW), chlorophyll b (98.70 mg g-1 FW) and total chlorophyll (135.70 mg g-1 FW) respectively, were recorded from the seedlings developed from GA3–(100 ppm)-primed seeds, whereas, in case of control minimum values were recorded for all the three chlorophyll fractions. Highest value for leaf nitrogen content (3.40%) was observed in seedlings that were developed from GA3-(100 ppm)-primed seeds followed by KNO3 50 ppm (3.25%), whilst the minimum value of 1.64% was recorded from control. Highest leaf phosphorus content was recorded in seedlings that were developed from GA3-(50 ppm)-treated seeds (0.26%) followed by GA3-(100 ppm)- (0.25%) and water soaked-seeds (0.23%), whilst the minimum value (0.13%) was recorded in control seedling. Leaf potassium content significantly varied in the developed seedlings due to the influential role of different seed priming chemicals, and the maximum value for leaf potassium content (3.42%) was recorded from GA3-(100 ppm)-primed seedlings, that was at par with results of GA3 50 ppm, KNO3 50 ppm, KNO3 100 ppm and Thiourea 100 ppm. The propitious effects of GA3 in regulating metabolic activities of photosynthetic pigments was reported by Afroz et al. (2005) in mustard. Application of GA3 in seed treatment ameliorated the primary growth potential, activity of carbonic anhydrase, and NPK use efficiency. As a consequence, the available nutrients in the growth medium might have been absorbed more rapidly due to their maximum utilization by the developing seedlings (Khan et al., 2009).

  • Conclusion

    GA3 (100 ppm) proved best seed priming treatment for wood apple as it helps in increasing germination percentage to an extent of 96.01% higher than control beside enhancing vegetative growth (85.21% more lengthy seedling than control), increasing leaf chlorophyll content, and leaf mineral composition in order to procure seedlings with vigorous growth.

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Sau S, Pal B, Sarkar S, Sarkar T. Influence of Seed Priming on Germination and Seedling Vigour of Wood Apple (Feronia limonia Swingle) IJBSM [Internet]. 18Apr.2019[cited 8Feb.2022];10(1):128-136. Available from:

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