Abstract: In the present study, we have tested the effect of humic acid on seed germination of Radish (Raphanus sativus). Seeds were soaked in various concentrations (0.1%, 0.25%, 0.5%, 0.75% and 1%) of humic acid at different time periods (10, 60, 120, 180 and 240 minutes). After 7 days, the seeds were analysed for their germination capacity, root and shoot length. The study infers that humic acid with the concentration of 0.25% showed maximum seed germination (100%) and the optimum shoot and root length was recorded as 6.175cm and 11.46cm respectively after 60 minutes soaking

Alien Maria Roniesha

Masilamani Selvam

M¹ Sai Nandhini R¹

Prakash P¹

Stanley Abraham L²

Thirugnanasambandam R

CiteSeerX

 
 
 
International Journal of ChemTech Research  
                                                                  CODEN (USA): IJCRGG      ISSN : 0974-4290 
                                                            Vol.6, No.9, pp 4180-4185, September  2014 
 
 
RTBCE 2014[12th August 2014]  
Recent Trends in Biotechnology and Chemical Engineering 
 
 
Effect of Humic Acid on Seed Germination of Raphanus 
sativus L. 
 
Prakash P¹*, Alien Maria Roniesha M¹, Sai Nandhini R¹, Masilamani 
Selvam M1, Thirugnanasambandam R.2  Stanley Abraham L²,  
 
1Dept of Biotechnology, Sathyabama University, Rajiv Gandhi road,  
Chennai-600119, Tamilnadu, India 
2Centre for Ocean Research, Sathyabama University, Rajiv Gandhi road,  
Chennai - 600 119, Tamilnadu, India 
        
*Corres.author: kpprakashmtech@gmail.com  
 
Abstract: In the present study, we have tested the effect of humic acid on seed germination of Radish 
(Raphanus sativus). Seeds were soaked in various concentrations (0.1%, 0.25%, 0.5%, 0.75% and 1%) of 
humic acid at different time periods (10, 60, 120, 180 and 240 minutes). After 7 days, the seeds were analysed 
for their germination capacity, root and shoot length. The study infers that humic acid with the concentration of 
0.25% showed maximum seed germination (100%) and the optimum shoot and root length was recorded as 
6.175cm and 11.46cm respectively after 60 minutes soaking. 
Key words: Humic acid, Raphanus sativus, seed germination, plant growth studies. 
 
Introduction 
Raphanus sativus belong to the brassicaceae family and is distributed all over the world. They vary in 
their size (10-15cm), colour (ranging from white or red) and in cultivation duration. It grows best in sandy 
loams and full sun light with pH 6.5-7.0. It is rich in ascorbic acid, folic acid and potassium. The root of R. 
sativus has several polyphenolics like vanillic acid, phenyl pyruic, syringic acid, p-coumaric, o-coumaric and 
caffeic
1,2
. Wild radish seed has 48% of oil content and used as biofuel. Radish are rich in potassium, which 
helps in maintaining the blood pressure and have benefits in certain ailments like jaundice, piles, urinary 
disorder, weight loss, constipation, cancer, skin disorder and insect bites
3
. In this study, effect of humic acid on 
seed germination of radish was evaluated. Humic acid are the component of humus, consist of humic, fulvic 
and hymatomelonic acid
4
, which are formed by organic decomposition of organic matter like lignites, peat, as 
well as from abiotic organic matter like soil and water ecosystem. Structure of humic acid complex found to be 
organic containing polyphenolic backbone with carbohydrate and peptide side chain. So, they have high 
molecular weight 
5&6
. In general, humic acid supplies both macro and micro nutrients to growing plants, 
increases soil fertility and productivity, increases water holding capacity and enhance seed germination. Humic 
acid also reduces the other chemical fertilizer requirements, increases aeration of the soil, increases the protein 
and mineral contents of most crops
7
. Hence the objective of this present study is to gain information on humic 
acid and its effect on seed germination of Raphanus sativus L. 
www.sphinxsai.com 
Prakash P et al /Int.J. ChemTech Res.2014,6(9),pp 4180-4185. 4181 
 
Materials and Methods 
Radish seeds were purchased from local market and healthy seeds were selected for this study. The 
seeds (10 Nos) were soaked in sterile distilled water and treated with various concentration of humic acid 
(0.1%, 0.25%, 0.5%, 0.75% and 1.0%) for various time intervals (10 min, 60 min, 120 min, 180 min and 240 
min). All the treated seeds were placed in sterile petridishes containing a thin layer of wet cotton and covered 
with wet filter paper. Seed germinated in distilled water was served as a control. The percentage of seed 
germination, root length and shoot length were recorded after 7 days of sowing and tabulated.  
Results and Discussion 
Table 1: Effect of Humic acid on seed germination (%) 
Seed soaking time 
( minutes) 
Humic acid concentration 
(%) 
Seed germination 
(%) 
10 0.1 90 
0.25 90 
0.5 80 
0.75 90 
1.0 90 
60 0.1 100 
0.25 100 
0.5 90 
0.75 80 
1.0 90 
120 0.1 70 
0.25 90 
0.5 90 
0.75 90 
1.0 90 
180 0.1 90 
0.25 90 
0.5 90 
0.75 90 
1.0 90 
240 0.1 90 
0.25 80 
0.5 80 
0.75 80 
1.0 80 
Control Soaked in Water 80 
 
In the present study, effect of humic acid treated radish seeds showed significant results over untreated 
seeds and are presented. (Table- 1) shows the percentage of seed germination over control in different 
concentrations of humic acid. Among the treatments, maximum seed germination was observed in 0.1% and 
0.25%. Similarly Patil et al
8
 reported that the application of 1% of potassium humate with deproteinized leaf 
extract increased the seed germination of soybean and black gram up to 100%.  Volenec et al
9
 stated that 
deproteinized juice (DPJ) consists of many free aminoacids, vitamins, hormones, and the foliar application of 
deproteinized juice significantly increased the growth and yield of crop plants. Radish seeds after germination 
were observed for their growth in terms of shoot and root length between 24 and 168 hrs and the results 
obtained were showing significant improvements than control seeds (Table- 2 & 3). The maximum shoot length 
was recorded as 6.175cm at 168 hrs in 0.25% of humic acid. The same trend was observed in root length i.e. 
11.4cm at 168 hrs in 0.25% of humic acid with 60 minutes treatment. Xue et al
10 
observed better performance 
when humic acid was applied along with chemical fertilizer in maize, wheat, cotton, rapeseed and sesame 
production as compared to Di-ammonium phosphate and other fertilizers. Humic acid increases resistance of 
Prakash P et al /Int.J. ChemTech Res.2014,6(9),pp 4180-4185. 4182 
 
the crop against drought, cold and diseases, resulting in overall improvement of crop yield and accumulation of 
availability to the plant nutrients. It induces and increases plant growth sustainably by releasing its nutrients 
slowly to the plants and microflora associated with it and expressed its effect on root enzymes and rhizosphere 
soil
11&12
. Trichoderma viride has effectively converted humic acid from Lignite and similar effect was obtained 
on Sorghum and Stevia rebaudiana and the same author also reported that 60% of potassium humate enhances 
the mushroom growth, 4% enhances Morus alba and 0.2% of humic acid enhancing the growth of Spirulina 
plantensis
13-17
. Kazuhiro Sugimoto
18
 reported that wild radish seeds A were capable of tolerating NaCl at 25 °C, 
with tolerance of 10 mM MgCl2 and 100 mM MgSO4, but was sensitive to NaCl and CaSO4. The growth 
response of the humic acid are due its hormonal effect and rich in macro and micro nutrients Atiyeh et al.
19
. 
Similar effect was found in this study and can be used effectively for seed treatment and high crop yield. 
Conclusion  
 Thus the present study reveals the required concentration of 0.25% of potassium humate for seed 
germination and seedling growth with effective root length and shoots length. This can be used for both seed 
treatment and growth regulator. 
 
Acknowledgement  
The authors are grateful to the management of Sathyabama University for providing laboratory 
facilities, support and encouragement. 
 
 
 
 
 
 
 
 
 
 
Prakash P et al /Int.J. ChemTech Res.2014,6(9),pp 4180-4185. 4183 
 
  
 
 
 
Root length (mm) 
  
Total  root 
length  (mm) 
Average root 
length (mm) 
Soaking 
Time 
(minutes) 
  1 2 3 4 5 6 7 8 9 10     
  Control 16.5 12.2 7.8 1.4 10.5 6.1 4.8 2.1 7 0 68.4 6.84 
10  
0.1 13.9 13.7 14.3 6.6 7.1 7.9 14 6.1 4 0 87.6  8.76 
0.25 9.8 5.4 6.1 4.2 2.1 2.4 2.1 3.4 9.5 0 45  4.5 
0.5 10 4.7 15.9 5.1 6.7 11.9 9.1 5 0 0 68.4  6.84 
0.75 12.4 8.6 11 0.5 1.8 11.6 6.7 15 9.6 0 77.2  7.72 
1 10.3 12.5 14.6 4.1 7.5 4.3 0.5 3.2 10.5 0 67.5  6.75 
60  
0.1 7.5 7 9.5 13 11 6 9 15 2.5 7 87.5  8.75 
0.25 11.3 14.8 16.5 12.8 13.3 8.1 12 13.2 2.3 10 114.55  11.46 
0.5 8.9 9 10.8 8.6 9.1 12 4.2 6 5 0 73.6  7.36 
0.75 2 9.2 5.3 4.2 7.4 9.4 1 4.1 4 0 46.6  4.66 
1 7.4 9.5 3.1 15.2 9.9 12.1 9 9.8 8.9 0 84.9  8.49 
120 
0.1 11.2 14.2 12.5 10 2 9.2 11 5 7 0 82.1  8.21 
0.25 11 10.5 11 7.5 6.7 6.2 11.6 9.7 2.9 0 77.1  7.71 
0.5 15 9 15 15.2 8.4 5.5 5.4 10.6 9.4 0 93.5  9.35 
0.75 14.5 14 7 4.2 6.5 10 8.5 2.5 4 0 71.2  7.12 
1 15.5 10.5 7 11.2 11.9 10.6 9.8 9 9.8 0 95.3  9.53 
180 
0.1 8.5 13.2 10.5 10.5 6 7.2 9 1.2 2.5 0 68.6  6.86 
0.25 6 8.3 10.9 7 11.2 11.5 7.8 1.2 9.3 0 73.2  7.32 
0.5 6.5 10.4 1.5 5.9 9.7 8.6 9.4 10.6 13.2 0 75.8  7.58 
0.75 0.7 1.2 10 14.4 12.2 5.4 5.1 12.2 3 0 64.2  6.42 
1 8.2 16.3 9.9 13.1 9 6.8 9.2 10 15.5 0 98  9.8 
240 
0.1 10.5 8 15 10.1 10 6.8 5.6 13.2 7.6 0 86.8  8.68 
0.25 13 11.5 9.5 7.9 5.8 5 9.1 11.3 0 0 73.1  7.31 
0.5 14 8.7 15 14.7 9.5 15.7 15 8.6 0 0 101.2  10.12 
0.75 14 12.5 14.5 3.7 14.3 8.9 6 12.3 0 0 86.2  8.62 
1 15 9.5 6 6.7 14.6 7.7 6.8 9.9 0 0 76.2  7.62 
Table 2: Average root length after treatment of different concentration of Humic acid 
 
 
Prakash P et al /Int.J. ChemTech Res.2014,6(9),pp 4180-4185. 4184 
 
  
 
 
 
Shoot length (mm) 
  
 
 
Total shoot 
length  
(mm) 
 
Average shoot 
length (mm) 
 No of Seeds  1 2 3 4 5 6 7 8 9 10     
time  control 7.6 6.9 5.6 4.5 4.1 2.4 1.9 2.6 3 0 38.6 3.86  
10 min 
0.1 6 4.7 6.6 2.3 6.3 5.4 5.2 3.4 3 0 43  4.3 
0.25 5 4.2 2.5 2.4 2.5 1.6 1.9 2 2.2 0 24.55  2.46 
0.5 4.5 5.4 8.4 4.2 4 4.7 6.2 3.1 4 0 45  4.5 
0.75 3.2 3.7 1.9 1.3 3.2 6.2 5.3 5.1 3.1 0 33.75  3.38 
1 6.4 4.1 3.9 3.8 5.3 3.2 1.5 3.4 5 0 37.6  3.76 
60 min 
0.1 4.1 4.5 2.8 4.5 3 2 1.5 4 4 5.2 35.7  3.57 
0.25 6.7 7 6.3 6 6 4.5 5 7 6 7 61.75  6.175 
0.5 3.2 4.1 5.3 3.8 3.7 3.8 3.6 4 4.1 0 36.1  3.61 
0.75 3.8 3.7 1 3.1 1.1 4.1 2.2 3 3.2 0 25.95  2.59 
1 5 4.2 4 5.9 5.5 5.3 5.8 5.5 6 0 48.2  4.82 
120 min 
0.1 3 3.2 5.5 1.2 1 3.5 3 4 3.3 0 27.8  2.78 
0.25 3.5 5.5 2 1.8 3.1 5.4 4.8 3.7 2.9 0 32.95  3.295 
0.5 5.2 3.9 5.7 4.3 4 4.2 3.2 3.8 1.5 0 36.3  3.63 
0.75 9.6 9.5 7.5 8.5 4 7 5.5 4.2 3.9 0 60.45  6.05 
1 4.5 5 6 4 6.5 3.7 6.2 4.2 5.5 0 46.6  4.66 
180 min 
0.1 2 4.5 5.5 1 2.5 5 1.7 2.5 2.4 0 27.2  2.72 
0.25 6.2 2 5 6 3.7 6.5 4.3 4.5 2.9 0 41.35  4.14 
0.5 3 2.5 1.4 2.8 2.1 2.8 1 4 3 0 23.1  2.31 
0.75 3.2 4.5 2.5 4.5 2.5 3 2.5 2.5 2 0 27.95  2.79 
1 1.5 5.4 4.6 5.4 3.9 4 3.2 4.1 3.9 0 37  3.7 
240 min 
  
  
  
  
0.1 5.5 5.5 5 4.1 3 4.9 4.7 4.8 3.3 0 40.9  4.09 
0.25 3.5 1.5 5.5 5.3 1.9 2.4 4 4.4 0 0 28.75  2.88 
0.5 6 5 4 3.7 4.7 3.6 5.1 7.1 0 0 39.7  3.97 
0.75 3 4.5 5 4.8 5.8 4 4.4 3.4 0 0 35.65  3.57 
1 5 4.2 4.4 4.2 5.1 3.6 4.9 2.8 0 0 35.2  3.52 
 
Table 3: Average shoot length after treatment of different concentration of Humic acid 
 
 
Prakash P et al /Int.J. ChemTech Res.2014,6(9),pp 4180-4185. 4185 
 
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Effect of Humic Acid on Seed Germination of Raphanus sativus L

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Effect of Humic Acid on Seed Germination of Raphanus sativus L

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