10 April 2025

INTRODUCTION :- Suran, also known as Elephant Foot Yam, is an important tuber crop valued for its nutritional and medicinal properties. Conventional propagation through corms is slow and limited due to low multiplication rate and high risk of disease transmission. Plant tissue culture offers an efficient alternative for mass multiplication of disease-free, high-yielding Suran plants.

EXPLANT CAN USE :-

corm buds

shoot tip

PROPAGATION OF SURAN :-

After discussion with Dixit Sir, the plan is to do plant tissue culture of suran by taking it from the market, cutting it into equal sizes, and planting it in the soil.

We bought 2 kg of suran from the market, cut it into 4 pieces, and planted them in the soil.

19 November 2025

After nine months of cultivation, Suran (Amorphophallus paeoniifolius) tubers weighing 2 kg at the time of planting produced a total yield of 10 kg at harvest.

21December 2025

STAGES :-

1.Preparation of Explant

2. Initiation

3. Shoot Multiplication

4. Rooting

5. Hardening/Acclimatization

Table No. 1 MATERIALS :-

Beaker Measuring cylinder
Test tubeDistilled water
Conical flaskCulture bottle
ScissorsTissue paper
Surgical sterile bladeForceps
Autoclave bagSterile Distilled water
ScalpelFilter paper
Test tube standSpatula
Petri plateCotton plug

CHEMICALS FOR STERILIZATION :-

1. Luquid soap

2. Dettol

3. Sodium hypochlorite

4. Antibiotics

CHEMICALS FOR MS MEDIA :-

1. Macronutrients

2. Micronutrients

3. Vitamins

4. Iron source

5. Sucrose

6. Inositol

7. Agar

8. HCl/ NaOH

Table No. 2 PREPARATION OF MACRONUTRIENTS :-

MacronutrientsOriginal ConcentrationAmount of salt after multiplying original concentration by 20X, Solution Volume (250ml)
NH4NO3 16508250mg
KNO319009500mg
KH2PO4170850mg
MgSO4.7H2O3701850mg
CaCl2.2H2O4402200mg

Table No. 3 PREPARATION OF MICRONUTRIENTS :-

MicronutrientsOriginal ConcentrationAmount of salt after multiplying original concentration by 100X, Solution Volume (250ml)
H3BO36.2155mg
MnSO4.4H2O22.3557.5mg
ZnSO4.7H2O8.6215mg
Na2MoO4.2H2O0.256.25mg
CoCl2.6H2O0.0250.625mg
CuSO4.5H2O0.0250.625mg
KI0.8320.75mg

Table No. 4 PREPARATION OF VITAMINS :-

VitaminsOriginal Concentration Original Concentration multiplying by 1000X, Final solution volume (50ml)
Thiamine HCl 0.15mg
Pyridoxin HCl0.525mg
Nicotinic0.525mg
Glycin2.0100mg

Table No. 5 PREPARATION OF IRON SOURCE :-

Iron sourceOriginal ConcentrationOriginal Concentration multiplying by 400X, Final solution volume (50ml)
FeSO4.7H2O27.82780mg
Na2EDTA.2H2O 37.33730mg

Table No. 6 PREPARATION OF MS MEDIA :-

MaterialsFinal Volume ( 250ml )
Macronutrients 12.5ml
Micronutrients0.625ml
Vitamins0.625ml
Iron source 0.250ml
Mesoinositol25mg
Sucrose7.5gm
Agar2gm
Distilled water 250ml

MESOINOSITOL:- When we prepare 1 litre ms media then we will take 100mg mesoinositol.

SUCROSE (3%):- When we prepare 1 litre ms media then we will take 30gm sucrose.PREPARATION OF

HORMONES:-

BAP Stock:– 100mg BAP in conical flask (250ml) dissolve with ethanol or 1M NaOH solution and BAP powder dissolve completely in solution and make final volume 100ml solution with Distilled water.

IBA Stock:- 100mg of IBA in conical flask (250) dissolve with ethanol or 1M NaOH solution and IBA powder dissolve completely in solution and make final volume to 100ml with Distilled water.

pH CHECK:- Prepare solution media pH range is 5.8 – 5.9 by using HCl or 1M NaOH.Agar:- Take 8 gm agar powder and add in media ,mix with glass rod then in microwave the media for 14 minutes and agar dissolve completely in media.

STERILIZATION PROCESS :-

Autoclave the used media to sterilize it before disposal. Discard the used media from the culture bottle. Soak the empty culture bottle in chromic acid (for cleaning or decontamination) for 24 hours. After 24 hours, remove the culture bottle from the chromic acid and wash it with liquid soap. Then dry it in a hot air oven at 180°C for 12 hours. Then we have wrapped glassware in autoclave bag then put in autoclave with distilled water and culture media.

Autoclave work at high pressure and temperature ( 15 psi, 121°C) for 15 to 20 minutes and kill the microorganisms. After sterilization removed glassware from sterilizer. And placed in laminar air flow then when we use it, we will turn8 on UV – ray light for 1 hour.

METHODOLOGY :-

21 December 2025

1.Select mother plants ( Shoot tips, Corm bud ) that are young shoot tips disease-free, and have high-yield and desirable traits.

2. Cut the mother plant in desirable shape and placed dry tray.

3. Wash the cut explant in running water for 15 minutes.

4. Handle the plant material with sterile gloves.

5. Then prepare total volume 300ml soap solution (30ml soap liquid and 270ml distilled water).6. Wear glove hand and wash the plant material for 5 minutes.

6. Then remove the soap solution and placed in glass beaker.

7. Cut the material desirable size with the help of knife.

8. Then soak the material in savlon solution ( 3ml savlon and 300 ml distilled water) for 15 minutes.

9. Then wash the plant material from sterile distilled water 3 to 5 time in bottle beaker.

11. Prepare 4% sodium hypochlorite solution and taking sodium hypochlorite solution into a laminar air flow and soak the plant material in sodium hypochlorite solution for 20 minutes( for 1 jar 10ml sodium hypochlorite and 250ml sterile distilled water).

12. After 20 minutes then remove plant material from sodium hypochlorite solution and wash the plant material 3 to 5 times in sterile distilled water.

13. Then cut the plant material required culture bottle size with the help of knife on sterilized brown paper.

14. Prepare antibiotic solution using Cefotaxime antibiotic( 1ml antibiotic and 200ml sterile distilled water) and soak the plant material in this solution for 30 minutes. Using only fresh antibiotic solution.

15. Then inoculate the plant material in culture medium( bottle beaker) and remove laminar air flow placed inoculate culture bottle at 24°C with photo periods of 6 to 7 hours light and 8 hours darkness for tissue culture sterile environment.

Result

Contamination appeared after 4 days.

24 December 2025

Again select a healthy suran plant and wash the explant ( shoot tip ) thoroughly. Sterilize it using a liquid soap solution, followed by soaking in an antibiotic solution for 30 minutes. After sterilization, inoculate the explant into a culture bottle under aseptic conditions.

Result

Contamination appears again after 4–5 days.

28 December 2025

I discussed the contamination issue of Suran with Dr. Dixit. He suggested preparing a 0.25% sodium hypochlorite solution and soaking the Suran in it for 30 minutes. After that, surface sterilization should be done, and the explants should be inoculated into MS medium.

Healthy yam (Suran) tubers are selected as explants and washed under running water for 15 minutes. Then, they are soaked in a 0.25% sodium hypochlorite solution for 30 minutes. After that, they are soaked in 200 ml of distilled water containing 2 g of Bavistin for 30 minutes. The tubers are then washed 3 to 4 times with distilled water. Finally, surface sterilization is done, and the explants are inoculated into MS medium.

29 December 2025

Healthy yam (Suran) tubers are selected as explants and washed under running water for 15 minutes. Then, they are soaked in a 0.25% sodium hypochlorite solution for 30 minutes. After that, they are soaked in 200 ml of distilled water containing 2 g of Bavistin for 30 minutes. The tubers are then washed 3 to 4 times with distilled water. Finally, surface sterilization is done, and the explants are inoculated into MS medium.

Result

The culture gets contaminated after one week.

6 january 2026

Suran is soaked in a sodium hypochlorite solution, then soaked in a Bavistin solution for one hour, followed by surface sterilization, and then inoculated onto MS medium.

Result :

suran got contaminated after one week.

11 January 2026

Suran is soaked in sodium hypochlorite solution, then soaked in Bavistin solution for one hour, followed by surface sterilization. The explants are then cut into 1 cm pieces and inoculated onto MS medium.

Result :

Suran showed contamination after one week.

14 January 2026

Suran is first soaked in sodium hypochlorite solution, then in Bavistin solution for one hour. After surface sterilization, the explants are cut into 1 cm pieces and placed on MS medium for inoculation.

Result :

After one week, contamination was observed in Suran.

16 January 2026

Suran is first cleaned with a sodium hypochlorite solution, then kept in a Bavistin solution for one hour. After sterilizing, the explants are cut into 1 cm pieces and placed on MS medium for growing.

23 January 2026

Suran is first soaked in sodium hypochlorite solution, then in Bavistin solution for one hour. After surface sterilization, the explants are cut into 1 cm pieces and placed on MS medium for inoculation.

Had a discussion with Dixit sir about suran. The cells were getting damaged due to the sodium hypochlorite solution, so sir suggested using monochloramine-T instead of sodium hypochlorite.

1 Fabruary 2026

First, Suran is soaked in Bavistin solution for 2 hours, then its surface is sterilized and inoculated into MS medium.

6 Fabruary 2026 12 Fabruary 2026

First, the suran is soaked in dilute sodium hypochlorite for 1 hour, then kept in Bavistin solution for 2 hours. After that, surface sterilization is done and it is inoculated in MS medium.

Discussion

Discussion was done with Dixit sir regarding contamination in the suran plant. He suggested that the suran should be washed continuously with UV-treated water for 12 hours. After that, it should be kept in Bavistin for 2 hours. Then surface sterilization should be done, and finally it should be inoculated onto MS medium.

27 February – 05 March 2026

Suran was washed continuously with UV-treated water for 12 hours. After that, it was kept in Bavistin for 2 hours. Then surface sterilization was done, and finally it was inoculated onto MS medium.

06 – 12 March 2026

Suran was washed continuously with UV-treated water for 12 hours. After that, it was kept in Bavistin for 2 hours. Then surface sterilization was done, and finally it was inoculated onto hormons medium.

13 – 19 March 2026

Suran was washed with clean UV- treated water for 12 hours. Then it was kept in bavistin for 2 hours. After that, it was sterilized and placed on hormone medium for growth.