Effect of Different Physical and Chemical Treatments on Seed Germination and Seedling Growth in Charoli (Buchanania lanzan Spreng.)

Introduction

Charoli (Buchanania lanzan Spreng.), which is also widely known as Chironji or Piyal, is a valuable fruit tree belonging to the Anacardiaceae family. The species originally originated from the Indian subcontinent. It was first documented by Francis Hamilton in 1798, and its valid scientific name was officially described in 1801 by the German botanist Kurt Sprengel (abbreviated as "Spreng." in the scientific name). Because of its immense ecological and economic value, this tree serves as an excellent option for social agroforestry, which integrates tree planting with local crops to benefit farming communities and support the environment. It assumes great significance due to its diverse uses and capacity to withstand adverse climatic conditions. At present, it grows under forest conditions as an underexploited fruit and provides monetary benefits to the tribal communities of the country. The kernels are highly nutritious, rich in protein (25-30%) and contain a sweet oil that can be used as a substitute for olive and almond oils. It is a medium-sized tree, reaching up to 40-50 feet in height with a straight trunk. The trees exhibit an alternate bearing habit, similar to mango. It flowers in the months of January and February and the fruits ripen in April and May. The kernels are of very high value and fetch Rs. 1800-2000 per kg in the market. In Vidarbha (Maharashtra), this species is found mainly in the Gadchiroli, Gondia, Amravati and Akola districts. Because charoli is a vulnerable plant, it is included in the Red Data Book published by the International Union for Conservation of Nature and Natural Resources (IUCN). This species has high socioeconomic value, providing a livelihood to the tribal population and holds high potential as a commercial horticultural crop. Unfortunately, overexploitation and indiscriminate harvesting (lopping and cutting) pose a severe threat to it; hence, urgent conservation efforts are required at all levels. Charoli seeds are recalcitrant in nature and lose viability in three months after harvesting (Chauhan et al., 2020) [2] . They exhibit poor germination even when exposed to favorable conditions due to exogenous seed dormancy. This dormancy may be attributed to morphological factors, such as a hard, thick testa, or to incorrect storage and handling (secondary dormancy). Such seeds require special physical treatments, such as stratification or scarification to overcome dormancy. Furthermore, pre-sowing seed treatments with chemicals like GA₃, KNO₃ and NAA (Rajamanickam et al., 2002) [14] have been reported to improve seed germination and seedling growth. To meet the local demand for quality charoli planting material, a field trial was conducted to find an effective seed treatment that facilitates the easy multiplication, regeneration and conservation of this important species. Fresh, fully ripened, uniform and disease-free charoli fruits were collected from the All India Coordinated Research Project (AICRP) for Dryland Agriculture, Dr. P.D.K.V., Akola. The seeds were manually extracted and washed thoroughly with water. Floating, nonviable and damaged seeds were carefully removed. The remaining viable seeds were shade-dried for one hour. Finally, healthy seeds of uniform size and weight were selected for the experiment.

Materials and Methods

Various physical treatments, such as scarification with a ladle, scarification with H₂SO₄ (5%) for 10 minutes and stratification at 4 0 C for 15 days, were applied to the freshly extracted charoli seeds. The seeds were then dipped in solutions of gibberellic acid (GA₃) (200 ppm and 300 ppm) and naphthaleneacetic acid (NAA) (200 ppm and 400 ppm) for 24 hours before sowing. After seed treatment, the seeds were shade-dried for 10 minutes and sown at a depth of 1 cm in 20 × 15 cm polythene bags. These bags were kept in a polyhouse. Bags contained a growth medium consisting of a 1:1:1 ratio of soil, farmyard manure (FYM) and sand. The bags were watered as needed. Observations for germination parameters were recorded daily, while vegetative parameters were recorded monthly, starting from the date of seed sowing.

Days required for germination

The date of first plumule emergence was observed. The time elapsed between the date of sowing and the date of plumule emergence was recorded as the number of days required for germination.

The germination percentage was recorded at 30 days after sowing (DAS). It was calculated using the following formula:

Germination percentage (%) = Number of seeds germinated Total number of seeds sown 𝑥 100

The survival percentage was recorded at 365 days after sowing (DAS). It was calculated using the following formula:

Survival percentage (%) = Number of seedlings survived Number of seeds germinated 𝑥 100

Number of leaves

The total number of leaves per plant was counted from ten randomly selected and tagged seedlings at 180 DAS. The mean value was computed and expressed as the average number of leaves per seedling.

The stem diameter of ten randomly selected and tagged seedlings was measured at a height of 3 cm from the base using a Vernier caliper at 180 DAS. The mean value was computed and expressed in millimeters (mm).

The height of ten randomly selected and tagged seedlings was measured from ground level to the growing tip at 180 DAS. The mean value was computed and expressed as the average seedling height in centimeters (cm).

At 180 days after sowing (DAS), ten randomly selected and tagged seedlings from each treatment were carefully uprooted and properly washed. The fresh weight of the entire plant (including both shoot and root systems) was measured immediately using an electronic balance. The average fresh plant weight was computed and expressed in grams (g).

Following the measurement of fresh weight, the same ten randomly selected and tagged seedlings were subjected to drying. The entire plants were dried in a hot air oven at 60-75 °C for 10 to 20 hours until a constant weight was achieved. The average dry plant weight was computed and expressed in grams (g).

The experimental data were subjected to statistical analysis using the analysis of variance (ANOVA) technique, as described by Panse and Sukhatme (1967) [11] .

Result and Discussion 1. Days required for germination

The days required for germination in charoli was significantly influenced by the different physical and chemical treatments and their combinations (Table 2, Fig. 1). Among the physical treatments, the minimum days to germination (13.67 days) was recorded in P 2 (Scarification with 5% H 2 SO 4 for 10 minutes), while the maximum (18.42 days) was observed in P 3 (Stratification at 4 0 C for 15 days).

Regarding the chemical treatments, C 2 (GA 3 300 ppm for 24 hours) recorded the minimum days to germination (12.44 days), whereas C 3 (NAA 200 ppm for 24 hours) recorded the maximum (18.11 days). Among the treatment combinations, the minimum days to germination (9.33 days) was achieved in T 6 (Scarification with H 2 SO 4 H 2 SO 4 for 10 minutes + GA 3 300 ppm for 24 hours), whereas the maximum (22.00 days) was recorded in T 11 (Stratification at 4 0 C for 15 days + NAA 200 ppm for 24 hours).

The reduction in the number of days to germination might be due to H₂SO₄ (5%) scarification, which degraded the lignin and cellulose matrix of the hard, stony endocarp. It plays a key role in breaking physical dormancy and increasing permeability for water imbibition and gaseous exchange. On the other hand, GA₃ (300 ppm) may be associated with triggering the de novo synthesis of hydrolytic enzymes like alpha-amylase and proteases, where it could rapidly mobilize stored endosperm reserves into simple sugars and amino acids and accelerate radicle emergence, which might be the reason for the reduced number of days to germination. Similar results were found by Sharma (2016) [16] , Joshi et al. (2017) [4] , Chauhan et al.

(2020) [2] , Sharma et al. (2020) [17] , Homeshvari et al. (2022) [3] and Prakash et al. (2025) [13] in charoli, Palepad et al.

(2017) [10] in custard apple and Lalitha et al. (2020) [6] in aonla. be associated with acting as a potent physiological catalyst to alter the endogenous promoter-to-inhibitor hormonal balance, where it could overcome Abscisic Acid (ABA) mediated suppression, promote cell division and elongation in the embryonic axis and ensure that a maximal number of embryos successfully break through the seed coat that might be the reason for the increased germination percentage. Similar results were found by Shukla and Solanki (2000) [18] , Narayan et al. (2014) [9] , Sharma (2016) [16] , Joshi et al.

(2017) [4] , Sadashiv et al. (2017) [15] , Ajith et al. (2018) [1] , Chauhan et al. (2020) [2] , Sharma et al. (2020) [17] , Thounaojam and Dhaduk (2020) [19] , Khobragade et al.

(2021) [5] , Homeshvari et al. (2022) [3] and Prakash et al.

(2025) [13] in charoli, Vasantha et al. (2014) [20] in tamarind and Palepad et al. (2017) [10] in custard apple.

The survival percentage of the seedling in charoli was significantly influenced by the different physical and chemical treatments and their combinations (Table 3, Fig. 2). Among the physical treatments, the highest survival percentage (67.25%) was recorded in P 1 (Scarification with ladle), while the lowest (54.17%) was observed in P 3 (Stratification at 4 0 C for 15 days). Regarding the chemical treatments, C 2 (GA 3 300 ppm for 24 hours) recorded the highest survival percentage (76.56%), whereas C 3 (NAA 200 ppm for 24 hours) recorded the lowest (53.56%). Among the treatment combinations, the maximum survival percentage (80.33%) was achieved in T 6 (Scarification with H 2 SO 4 5% for 10 minutes + GA 3 300 ppm for 24 hours), whereas the minimum (40.67%) was recorded in T 11 (Stratification at 4 0 C for 15 days+ NAA 200 ppm for 24 hours).

The increase in survival percentage might be due to H₂SO₄ (5%) scarification ensured that the emerging hypocotyl did not exhaust its energy reserves struggling against a tough endocarp. It plays a key role in preserving cellular energy for immediate vegetative growth and providing early, robust and uniform germination for a competitive head start. On the other hand, GA₃ (300 ppm) may be associated with enhancing early cell elongation and metabolic activity in the hypocotyl and primary roots, where it could lead to the rapid establishment of a functional root system, efficient nutrient and water uptake and fortification against early-stage damping-off or environmental stress that might be the reason for the increased survival percentage. Similar results were found by Narayan et al. (2014) [9] , Joshi et al. (2017) [4] , Chauhan et al. (2020) [2] , Sharma et al. (2020) [17] , Homeshvari et al. (2022) [3] and Prakash et al. (2025) [13] in charoli, Manekar et al. (2011) [8] and Lalitha et al. (2020) [6] in aonla, Patel et al. (2017) [12] in mango and Palepad et al.

(2017) [10] in custard apple.

Number of Leaves

The number of leaves in charoli seedlings was significantly influenced by the different physical and chemical treatments and their combinations (Table 3, Fig. 3). Among the physical treatments, the highest number of leaves (12.83) was recorded in P 1 (Scarification with ladle), while the lowest (10.50) was observed in P 3 (Stratification at 4 0 C for 15 days). Regarding the chemical treatments, C 2 (GA 3 300 ppm for 24 hours) recorded the highest number of leaves (14.56), whereas C 3 (NAA 200 ppm for 24 hours) recorded the lowest (10.44). Among the treatment combinations, the maximum number of leaves (15.67) was achieved in T 6 (Scarification with H 2 SO 4 5% for 10 minutes + GA 3 300 ppm for 24 hours), whereas the minimum (7.67) was recorded in T 11 (Stratification at 4 0 C 15 days + NAA 200 ppm for 24 hours). The increase in the number of leaves might be due to H₂SO₄ (5%) scarification, which broke the mechanical seed barrier early. It plays a key role in transitioning the plant from heterotrophic to autotrophic growth much sooner. On the other hand, GA₃ (300 ppm) may be associated with stimulating cell division and elongation in the apical meristem, where it could lead to faster internodal development, earlier differentiation of leaf primordia, and enhanced nitrogen metabolism and chlorophyll synthesis that might be the reason for the increased number of leaves. Similar results were found by Sharma (2016) [16] , Joshi et al.

(2017) [4] , Chauhan et al. (2020) [2] , Sharma et al. (2020) [17] , Mahadev et al. (2021) [7] , Homeshvari et al. (2022) [3] and Prakash et al. (2025) [13] in charoli, Vasantha et al. (2014) [20] in tamarind, Palepad et al. (2017) [10] in custard apple and Lalitha et al. (2020) [6] in aonla.

The stem diameter of the seedling in charoli was significantly influenced by the different physical and chemical treatments and their combinations (Table 4, Fig. 4). Among the physical treatments, the highest stem diameter (3.80 mm) was recorded in P 1 (Scarification with ladle), while the lowest (3.22 mm) was observed in P 3 (Stratification at 0 C for 15 days). Regarding the chemical treatments, C 2 (GA 3 300 ppm for 24 hours) recorded the highest stem diameter (4.11 mm), whereas C 4 (NAA 400 ppm for 24 hours) recorded the lowest (3.19 mm). Among the treatment combinations, the maximum stem diameter (4.47 mm) was achieved in T 6 (Scarification with H 2 SO 4 5% for 10 minutes + GA 3 300 ppm for 24 hours), whereas the minimum (2.80 mm) was recorded in T 11 (Stratification at 4 0 C for 15 days + NAA 200 ppm for 24 hours). The increase in stem diameter might be due to H₂SO₄ (5%) scarification, which facilitated rapid germination and early establishment. It plays a key role in allowing the root system to uptake optimal soil nutrients, which structures the stem. On the other hand, GA₃ (300 ppm) may be associated with enhanced cambial activity, where it could develop a larger initial leaf area, generate abundant photo-assimilates and promote radial cell division in the vascular cambium that might be the reason for increased stem diameter. Similar results were found by Sharma (2016) [16] , Joshi et al. (2017) [4] , Chauhan et al. (2020) [2] , Sharma et al. (2020) [17] and Mahadev et al. (2021) [7] in charoli, Vasantha et al. (2014) [20] in tamarind, Palepad et al. (2017) [10] in custard apple and Lalitha et al. (2020) [6] in aonla.

The height of the seedling in charoli was significantly influenced by the different physical and chemical treatments and their combinations (Table 4, Fig. 4). Among the physical treatments, the highest height of the seedling (34.11 cm) was recorded in P 2 (Scarification with 5% H 2 SO 4 for 10 minutes), while the lowest (26.70 cm) was observed in P 3 (Stratification at 4 0 C for 15 days). Regarding the chemical treatments, C 2 (GA 3 300 ppm for 24 hours) recorded the highest height of the seedling (36.09 cm), whereas C 3 (NAA 200 ppm for 24 hours) recorded the lowest (24.66 cm). Among the treatment combinations, the maximum height of the seedling (38.83 cm) was achieved in T 6 (Scarification with H 2 SO 4 5% for 10 minutes + GA 3 300 ppm for 24 hours), whereas the minimum (18.67 cm) was recorded in T 11 (Stratification at 4 0 C for 15 days + NAA 200 ppm for 24 hours).

The increase in seedling height might be due to H₂SO₄ scarification enabled early and uniform emergence. It plays a key role in giving the seedling a longer total duration for active vegetative growth during the season. On the other hand, GA₃ may be associated with the alteration of plant cell wall plasticity, where it could drive extensive cellular elongation in the stem internodes and enhance the sink strength of the apical meristem that might be the reason for increased seedling height. Similar results were found by Sharma (2016) [16] , Joshi et al. (2017) [4] , Sadashiv et al.

(2017) [15] , Chauhan et al. (2020) [2] , Sharma et al. (2020) [17] , Khobragade et al. (2021) [5] , Mahadev et al. (2021) [7] , Homeshvari et al. (2022) [3] and Prakash et al. (2025) [13] in charoli, Vasantha et al. (2014) [20] in tamarind, Palepad et al.

(2017) [10] in custard apple, Patel et al. (2017) [12] in mango and Lalitha et al. (2020) [6] in aonla.

The fresh weight of the seedling in charoli was significantly influenced by the different physical and chemical treatments and their combinations (Table 5, Fig. 5). Among the physical treatments, the highest fresh weight (20.27 g) was recorded in P 2 (Scarification with 5% H 2 SO 4 for 10 minutes), while the lowest (17.25 g) was observed in P 3 (Stratification at 4 0 C for 15 days). Regarding the chemical treatments, C 2 (GA 3 300 ppm for 24 hours) recorded the highest fresh weight (22.27 g), whereas C 3 (NAA 200 ppm for 24 hours) recorded the lowest (15.41 g). Among the treatment combinations, the maximum fresh weight (23.67 g) was achieved in T 6 (Scarification with H 2 SO 4 5% for 10 minutes + GA 3 300 ppm for 24 hours), whereas the minimum (14.07 g) was recorded in T 11 (Stratification at 4 0 C for 15 days + NAA 200 ppm for 24 hours). The increase in fresh weight might be due to H₂SO₄ scarification, which facilitated unhindered radicle emergence and the early development of a sprawling root system. It plays a key role in improving water and mineral uptake. On the other hand, GA₃ may be associated with increasing cellular volume and tissue hydration, where it could enhance overall morphometric traits such as plant height, girth and leaf area that might be the reason for increased fresh weight. Similar results were found by Sharma (2016) [16] , Mahadev et al. (2021) [7] , Homeshvari et al. (2022) [3] and Prakash et al. (2025) [13] in charoli, Vasantha et al. (2014) [20] in tamarind, Palepad et al. (2017) [10] in custard apple and Lalitha et al. (2020) [6] in aonla.

The dry weight of the plant in charoli was significantly influenced by the different physical and chemical treatments and their combinations (Table 5, Fig. 5). Among the physical treatments, the highest dry weight (10.08 g) was recorded in P 2 (Scarification with 5% H 2 SO 4 for 10 minutes), while the lowest (7.08 g) was observed in P 3 (Stratification at 4 0 C for 15 days). Regarding the chemical treatments, C 2 (GA 3 300 ppm for 24 hours) recorded the highest dry weight (12.09 g), whereas C 3 (NAA 200 ppm for 24 hours) recorded the lowest (5.21 g). Among the treatment combinations, the maximum dry weight (13.47 g) was achieved in T 6 (Scarification with H 2 SO 4 5% for 10 minutes + GA 3 300 ppm for 24 hours), whereas the minimum (3.87 g) was recorded in T 11 (Stratification at 4 0 C for 15 days+ NAA 200 ppm for 24 hours). The increase in dry weight might be due to H₂SO₄ (5%) scarification, which caused an early physical breach of the seed coat. It plays a key role in facilitating an advanced canopy structure for optimum light interception. On the other hand, GA₃ may be associated with the stimulation of cell division, cell elongation and auxin metabolism, where it could maximize the net assimilation rate, drive high rates of carbon fixation and optimize the source-sink dynamic that might be the reason for increased dry weight. Similar results were found by Sharma (2016) [16] and Homeshvari et al.

(2022) [3] in charoli, Vasantha et al. (2014) [20] in tamarind, Palepad et al. (2017) [10] in custard apple, Patel et al. (2017) [12] in mango and Lalitha et al. (2020) [6] in aonla.

Conclusions

Based on the results of the present investigation entitled "Effect of Different Physical and Chemical Treatments on Seed Germination and Seedling Growth in Charoli (Buchanania lanzan Spreng.)", It can be concluded that the seed treatment with 5% H₂SO₄ for 10 minutes followed by soaking in GA₃ at 300 ppm for 24 hours gave the earliest germination, the maximum germination percentage, optimum vegetative growth and the highest survival percentage of charoli seedlings.

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