Indian Journal of Agricultural Research

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Indian Journal of Agricultural Research, volume 57 issue 5 (october 2023) : 671-675

Germination Behavior of Rauvolfia serpentine (L.) Seeds with the use of Different Seed Treatments

Amit Kumar Tiwari1, Prawal P.S. Verma2, Dipender Kumar1,*, Sonveer Singh1, Rakesh Kumar3, R.C. Padalia1, R.K. Upadhyay1
1CSIR-Central Institute of Medicinal and Aromatic Plants, Research Centre, Pantnagar-263 149, Uttarakhand, India.
2CSIR-Central Institute of Medicinal and Aromatic Plants Research Centre, Purara, Post-Gagrigole, Bageshwar-263 641, Uttarakhand, India.
3CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow-226 015, Uttar Pradesh, India.
Cite article:- Tiwari Kumar Amit, Verma P.S. Prawal, Kumar Dipender, Singh Sonveer, Kumar Rakesh, Padalia R.C., Upadhyay R.K. (2023). Germination Behavior of Rauvolfia serpentine (L.) Seeds with the use of Different Seed Treatments . Indian Journal of Agricultural Research. 57(5): 671-675. doi: 10.18805/IJARe.A-5624.
Background: Sarpagandha is an indigenous medicinal herb of Indian continent. It has many medicinal properties. Due to increasing demand of Sarpagandha, its exploitation is increasing continuously but for lack of cultivation, this plant has been listed in endangered category. Commercially, it is propagated through seeds but the main barrier of seed propagation is its irregular and low germination. Hence, the current study was undertaken to study the effect of growing media and seed treatment methods on seed germination and seedling growth of Sarpagandha to eliminate the inhibitory effect of the hard stony endocarp.

Methods: Seed germination and seedling growth experiments of Sarpagandha were carried out with following different seed treatments, T1 (Control), T2 (Water soaking for 24 hours), T3 (water soaking for 48 hours), T4 (Water soaking for 24 hours + cotton cloth wrapping for 24 hours), T5 (GA3 200 ppm), T6 (FYM treatment for 24 hours), T7 (FYM treatment for 48 hours), T8 (Hot water treatment) and T9 (Cow dung treatment for 24 hours). Experiment was laid out in CRD. The data recorded on the various parameters were analyzed at 5% level of significance by using ANOVA.

Result: Results indicated highest germination percentage (51), lowest mortality percent (49), minimum days for initiation of germination (23), minimum days taken for germination completion (33), highest speed of germination (3.77), maximum vigour index (1441.44), highest root (11.27 cm) and highest shoot length (17 cm) in T5 (GA3 200 ppm) followed by T3 (water soaking for 48 hours). Indicating, T5 (GA3 200 ppm) to be best seed treatment for higher germination and subsequent seedling growth of Sarpagandha (Rauvolfia serpentine L.).
Sarpagandha [Rauvolfia serpentine (L). Benth. Ex Kurz] is one of the important medicinal plant of India, belong to family Apocynaceae. Sarpagandha is an indigenous medicinal herb of Indian continent, mentioned in ancient Indian medicinal literature, 3000 year back. “Sarpagandha” is Sanskrit word means smelling like snake which refers to the herb strong odor. The herb has a number of therapeutic benefits and has been used in Ayurvedic medicine for centuries. It’s also known as Chandrabhaga, Chandrika and Chota-Chand in Hindi, Shivanabhi balli Sarpagandhi, Guruda paatala and Paatala Gandhi in Kannada and Serpentine root, Serpentine wood and Indian snakeroot in English (Anonymous, 1956). Sarpagandha is a central nervous system stimulant and also an anti-hypertensive. Sarpagandha roots are known to treat intestinal disorders and also stimulate contractions. A number of different species of the Sarpagandha are found all over India. It is also known for its antimicrobial, antifungal, anti-inflammatory, antiproliferative, antidiuretic and anticholinergic activities. The plant is known for curing various disorders because of the presence of alkaloids (ajmaline, ajmalicine, ajmalimine, deserpidine, indobine, indobinine, reserpine, reserpiline, rescinnamine, rescinnamidine, serpentine, serpentinine and yohimbine carbohydrates), flavonoids, glycosides, phlobatannins, phenols, resins, saponins sterols, tannins and terpenes. The plant parts, root and rhizome have been used for curing a large number of diseases such as high blood pressure, mental agitation, epilepsy, traumas, anxiety, excitement, schizophrenia, sedative insomnia and insanity etc (Kumari et al. 2013). Due to the increasing demand of Sarpagandha, its exploitation is increasing continuously, due to lack of cultivation; the situation has reached to the extent that this plant has been listed in endangered category by International Union for conservation of Nature and Natural Resources (Jain et al. 2003). Since 1969, the Indian government has also prohibited the collection from forests and export of Sarpagandha. In order to meet the demand, it has become necessary to promote the cultivation of Sarpagandha and this is possible only when scientific cultivation is done at commercial level. In commercial cultivation, it is propagated through seeds. Irregular and low per cent germination is the main barrier of seed propagation of the Sarpagandha plant. Variation is found in the germination percentage of Sarpagandha seeds which is 10 to 40 per cent (Farooqui et al. 2001). This is partly associated with the adverse effects of the stony endocarp. The absence of embryo is another serious factor and is probably due to parthenocarpy or sterility. In relevant to the above fact for seed germination problem, aims to study the effect of growing media and seed treatment on seed germination and seedling growth of Sarpagandha such that the inhibitory effect of the hard stony endocarp can be eliminated.
The experiment was carried out at CSIR-Central Institute of Medicinal and Aromatic Plants Research centre Pantnagar, Udham Singh Nagar, Uttarakhand, during March to May in 2020. The experimental site is located at latitude of 29.00°N, longitude of 79.38°E and at an altitude of 243.84 m. It experiences climate of hot summer and chilled winter. The maximum temperature ranges between 35-45°C and the minimum between 2-5°C with average rainfall of 1350 mm. The soil was clay loam with neutral reaction (pH-7.1). The experiment was laid out in a Completely Randomized Design (CRD) with 9 treatment combinations and 3 replications. The experiment was comprised of following seed treatments T1 (Control), T2 (Water soaking for 24 hours), T3 (water soaking for 48 hours), T4 (Water soaking for 24 hours + cotton cloth wrapping for 24 hours), T5 (GA3 200 ppm), T6 (FYM treatment for 24 hours), T7 (FYM treatment for 48 hours), T8 (Hot water treatment) and T9 (Cow dung treatment for 24 hours). CIM-Sheel variety of Sarpagandha was collected from Research Farm of CIMAP Research Centre, Pantnagar, US Nagar, Uttarakhand. There was no treatment of seeds for T1 (Control), seeds were sown directly in poly bags. For T2, the seeds were soaked in water for 24 hours. The seeds were soaked in water for 48 hours for T3. For T4, the seeds were first soaked in water for 24 hours, then the seeds were removed and wrapped in moist cotton cloth for 24 hours, moisture was also maintained in the cotton cloth for 24 hours. For T5 treatment, the seeds were soaked in 200 ppm of GA3 solution for 24 hours. For T6 and T7, the seeds were kept in farm yard manure for 24 and 48 hours respectively. Seeds were soaked in a hot water of 80ºC for 24 hours in T8 (hot water treatment). For T9, the seeds were kept in cow dung for 24 hours. The seeds were sown directly in poly bags after seed treatment in all treatments; each seed was sown in separate poly bags at about 1cm depth. 200 seeds were taken from each treatment. Seed sowing was done in first week of March, 2020. Irrigation was applied immediately after seed sowing and later on irrigated on alternate days till the final emergence. 20 seedlings were studied for growth parameters. Data on germination percentage was recorded from the first germination until further germination at two days interval. The germination percentage was calculated by number of germinated seed divided by the total number of seeds and multiplied by 100. At the beginning of germination it was considered as days taken for initiation of germination. Days taken for completion of germination were calculated as the difference between initial and final emergence. Root length (cm) was measured by destructive method of uprooting the plants and taking measurement by standard method. Shoot length (cm) was measured when the seedlings were fully grown. Speed of germination and vigour index were calculated by following formula (Agrawal, 1995):
 
       
Where
n = number of germinated seeds, d= number of days.
 

The data recorded on the various characteristic of seed germination and seedling growth were subjected to Fisher’s method of ANOVA and interpretation of data was taken as per Sukhatme and Amble (1995).
Based on the analyzed statistical data presented in Tables 1 and 2, it was revealed that various treatments significantly affected the seed germination in Sarpagandha (Rauvolfia serpentine L). Based on the results obtained, it was found that all the treatments (T2 to T9) were better than control treatment. Results also indicated superiority of T5 (GA3 @ 200 ppm) for all the observed parameters, followed by T3 (water soaking for 48 hours). All the observations are discussed in detail below.
 
Germination percentage
 
Data pertaining to germination percent is presented in Table 1.

Table 1: Effect of seed treatments on germination percentage, mortality percent, days taken for initiation of germination and days taken for completion of germination of seeds of Sarpagandha (Rauvolfia serpentine (L).



Seed treated with GA3 @ 200 ppm (T5) recorded highest germination percentage (51) followed by T3 (water soaking for 48 hours) which was 45.33. Treatments T2 (Water soaking for 24 hours) (39.33) and T4 (Water soaking for 24 hours + cotton cloth wrapping for 24 hours) (39.67) were statistically at par with each other, respectively. The result of T6 (FYM treatment for 24 hours), T7 (FYM treatment for 48 hours) and T9 (Cow dung treatment for 24 hours) were 37.33, 32.33 and 34.67 respectively. Whereas T8 (hot water treatment) recorded the lowest germination percentage (32) as compared to other treatments. The result of control was found better than T7 and T9. Similar results were found by Bhuyar et al., (2000) and Ponkumar et al., (2008) in Rauvolfia serpentina, Mithra and Ghosh (2004) found similar result in Withania somnifera. Hussain and Jha (2014) also reported that germination percentage (56.66) of treated Rauvolfia tetraphylla seeds increased with the use of GA3, while it was 31.26 per cent in untreated seeds.
 
Mortality percent
 
Data pertaining to mortality per cent is presented in Table 1. Seeds not having sprouts were counted and the converted into mortality percentage. Results showed significant difference in treatments for mortality percentage. Significantly lowest mortality (49 per cent) was recorded in T5 (GA3 @ 200 ppm) compared to other treatments. It was followed by water soaking treatment for 48 hours (54.67%). The maximum mortality (68%) was found in T8 (hot water treatment). The results are in line with the findings of Afshan Niyaz (2014) who reported that the mortality per cent of W. somnifera can be reduced by pre-treating seeds with GA3 500µg/l for 24 hours before sowing, this increased germination percentage.
 
Days taken for initiation of germination
 
Data presented in Table 1. Compared to the all treatments, T5 (GA3 @ 200 ppm) recorded earliest germination (23 days). The results of T3 (water soaking for 48 hours treatment), T2 (water soaking for 24 hours treatment) and T4 (water soaking for 24 hours + cotton cloth wrapping for 24 hours treatment) is slightly less than T5, which was 25, 24 and 24 days respectively. Where as, maximum number of days taken for initiation of germination (35 days) was recorded in control. Chacko and Singh (1966) confirmed the findings who reported the higher rate of germination in papaya seeds treated with GA3. These results were also similar to the findings of Babu et al., (2010) in papaya.
 
Days taken for completion of germination
 
Data presented in Table 1. From the very first day when the germination begun till the completion of germination, number of days were counted. Significant difference was found in all treatments. Minimum number of days for completion of germination (33 days) was recorded in GA3 @ 200 ppm (T5)  at par with the T(water soaking for 48 hours treatment) and T4 (water soaking for 24 hours + cotton cloth wrapping for 24 hours treatment) which was 40 and 48 days. Whereas, control took maximum number of days for completion of germination (60 days). These results are in line with the findings of Anjanawe et al., (2013) who reported minimum number of days for completion of germination with GA3 treatment in papaya.
 
Speed of germination
 
Data pertaining to germination speed represented in Table 2. The vigour of seed is determined by high speed of germination. Significant differences were found in all treatments. The speed of germination was maximum in GA3 @ 200 ppm (3.77) followed by water soaking for 48 hours (2.93). The minimum speed of germination was found in T7 (1.53). Phatak et al., (2018) were also reported higher speed of germination (2.47) with the application of 1000 mg/l GA3 in sarpagandha.

Table 2: Effect of seed treatments on speed of germination, vigour index, root length (cm) and shoot length (cm) of seeds of Sarpagandha (Rauvolfia serpentine (L).


 
Vigour index
 
The mean data on vigour index is presented in Table 2. Vigour index was significantly influenced by the varied seed treatments. Highest vigour index (1441.44) was recorded with GA3 @ 200 ppm which was followed by T3 (1199.48). Lowest vigour index (671.5) was obtained with the T8 (hot water treatment). Parab et al., (2017) were also recorded highest vigour index with the application of GA3 @ 200 ppm in papaya.
 
Root length (cm)
 
The data on root length (cm) is presented in Table 2. The data revealed that the GA3 @ 200 ppm increased root length (cm) progressively. T5 i.e. GA3 @ 200 ppm recorded significantly longest root length (11.27 cm) closely followed by T3 i.e. water soaking for 48 hours (10.43 cm). Seed without treatment recorded shortest root length (8.17 cm) in data. The similar results were recorded by the Ponkumar et al., (2008) and Muneshwar (2015) in Sarpagandha.
 
Shoot length (cm)
 
The data on shoot length (cm) is presented in Table 2. The data revealed that the GA3 @ 200 ppm increased shoot length (cm) progressively. T5 i.e. GA3 @ 200 ppm recorded significantly longest shoot length (17 cm) which is at par with T3 i.e. water soaking for 48 hours (16 cm). Seed without treatment recorded shortest shoot length (11.5 cm). Velmurugan et al., (2003) also found and reported higher shoot length in ashwagandha crop after application of GA3.

All parameters were highly influenced by different treatments especially by GA3 @ 200 ppm, which is a growth hormone. The better performance of all parameters in Gibberellic acid treatment might be because Gibberellic acid is a plant hormone that helps the seed to germinate at a faster rate. The embryo is found inside the seed, which completes its life cycle under favorable conditions. Breaking of seed dormancy for the germination is controlled by some physical factors such as light, temperature and moisture and by the endogenous growth-regulating hormones such as Gibberellic acid (Hu et al., 2008; Rieu et al., 2008; Nester et al., 1988). Gibberellic acid stimulates seed germination. Gibberellic acid exerts its influence in two manners, first by increasing the growth potential of the embryo and second by inducing hydrolytic enzymes (Goto et al., 1999). During seed germination, embryonic gibberellic acid is released that triggers the weakening of seed cover by stimulating gene expression involved in cell expansion and modification. Gibberellic acid represents a natural regulator of the processes involved in seed germination to stimulate the production of hydrolytic enzyme (Koomneef and Vander Veen, 1980).
After the research findings, it is found and concluded that the application of Gibberellic acid (GA3) @ 200 ppm was the best among all treatments. Data proves that the performance of untreated seeds was significantly lower as compared to the other treatments. Lower germination of seed is a big problem in Sarpagandha cultivation. To overcome this problem Gibberellic acid (GA3) @ 200 ppm should be used for the highest seed germination and seedling growth in sarpagandha.
The authors are thankful to the Director, CSIR-CIMAP, Lucknow for providing facilities and cooperation during the period of investigation. 

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