In vitro STUDIES IN Baliospermummontanum (Willd) Mull. Arg.

Midhun M NairSchool of Environmental Sciences

MG University,Kottayam,Kerala,India

INTRODUCTION• Biotechnology is an advanced branch of biology

having many applications in different field of Biology such as agriculture, medicine, forestry, food science, environmental science etc

• Plant cell and tissue culture is one at the potential aspects of biotechnology. Plant tissue culture is the technique of growing plant cells, tissue and organ in an artificially prepared nutrient medium under aseptic conditions

MICROPROPOGATION

The most popular application of plant tissue culture is micropropagation which is an alternation to vegetative plant propagation.

1.Propagation using shoot tip/nodal segment culture2.Propagation by direct shoot organogenesion3.Propagation by indirect shoot organogenesis4.Propagation through somatic embryogenesis

HABIT

Baliospermum montanum (Willd) Mull. Arg. 

Baliospermum montanum Baliospermum montanum is a stout monoecious shrub, 1-

2m in height, branchlets cylindrical with leaves alternate. Flowers are small numerous, unisexual in axillary racemes

with male flower above, female flower below. The flowers are minute , about 3mm across, greenish

yellow in colour arranged in axillary and terminal racemes, spike or fascicles.

Seeds egg shaped,flowering and fruiting takes place from November to February.

Distribute throughout the most hilly regions from sub Himalayan tract to Deccan peninsula fairly common as undergrowth in the dismoist forests of Karnataka and Kerala.

It is used as a medical shrub.

SCOPE OF WORK Baliospermum montanum is an important medicinal plant

which become endangered due to the pharmaceutical over exploitation.

Roots of Baliospermum montanum is used to treat jaundice, anemiaetc .

The roots are sold under the name Dantimool or chotidanti. The drugs forms an important con-stituents of preparation like

Dantyaristam, Kaisoragulgula etc . For getting large quantity of roots the entire plant is destroyed

or disturbed. Micro propagation techniques have often been successful in

plant propagation. The quick multiplication is essential due to high economic

demand of the genus. Become constraints invitro technique can be adopted.

OBJECTIVES OF THE WORK• Axillary bud proliferation from

nodal explants• Callus induction• Shoot tip culture for shoot

elongation

MATERIALS AND METHODS• Several methods have been adapted for the in vitro culture at

explants tissue. • The general protocol for the plant tissue culture is as follows.1. Aseptic technique a) Sterilizing the culture vessels and instrumentsb) Preparation and sterilization of nutrient medium c) Sterilization of explant

2. Inoculation of explants3. Incubation

RESULTS

In the present study shoot tip, nodal segment, stem segments were taken as the explants.

Explants were inoculated and cultured on MS medium with different concentration and combination at hormones.

In different condition explants show different response.

Shoot tip

Shoot tip were inoculated on the MS medium containing hormones in different concentration and combination.

Shoot tips were cultured in BA alone and along with, IAA or IBA showered different response.

Maximum shoot tip elongation from shoot tip explants only in combination at BA(3mg/l) and IBA(.4 mg/l)

Nodal segment• Nodes were cultured in different in different

concentration and combination of BA and Kin alone or along with NAA, IBA, IAA and 2, 4-D.

• Medium containing BA along with IBA (0.1 mg/l) from nodal region low leaf axillary bud development and callus formation was observed.

• Concentration of IAA and BA was increased high degree of axillary bud proliferation was observed.

StemStem was cultured on MS medium containing

BA (2mg/l) along with IBA (5mg/l) showed moderate callusing.

When stem was cultured BA (5mg/l) along with 2,4-D (5mg/l) in high concentration, stem shows maximum rate of callus formation.

Callus sub culturing • Callus obtained from stem segments callus

on MS medium. IAA (2mg/l) along with KIN (4mg/l) rooting was obtained.

• Callus cultured on MS medium containing KIN (3mg/l) alone or KIN (2mg/l) along with BA (3mg/l) recallusing was obtained.

 Explants

 

 IAA

 IBA

 2,4-D

 BA

 Response

  

Shoot tip  

       0.1mg/

l

No response

   0.4mg/l

   3mg/l

Shoot tip elongation

  

Nodal segment

   

 0.5mg/l

      Low rate axillary bud elongation

 0.5mg/l

     4mg/l

Maximum rate axillary bud elongation

  

Stem   

  

 5mg/l

 

   2mg/l

Moderate callusing (++)

     5mg/l

 

 5mg/l

 

High callusing(+++)

 RESPONSE TO VARIOUS HORMONES

DISCUSSION The present work was undertaken to induce regeneration in

Baliospermum montanum (Willd.) Mull. Arg. Even though reports are available ininvitro propagation of

explants in Euphorbaceae.Shoot tip culture Maximum elongation of shoot tip was observed in BA(3mg/l)and

IBA(0.4mg/l) containing medium. BA has been reported to be a good hormone for regeneration in many plants.

KIN and NAA (0.2mg/l),no shoot formation. Only callusing was seen. This high callusing is due to the

presence of NAA. Auxins are known to promote high callusing and several cases

can let antogonostic to cytokinin.

Nodal segment culture KIN (4mg/l)and IAA(0.5mg/l) containing

medium show maximum axillary bud elongation.

IAA(0.5mg/l) alone containing medium and low rate of shoot tip elongation/axillary elongation.

Even though axillury bud proliferation in culture have been reported mostly in cytokinincontainina medium.

In the present study an auxin along with cytokinin was needed for maximum shoot multiplication .

CALLUS INDUCTION For stem segments moderate rate of

callusing was observed. Where stem was cultured in MS medium

containing BA (2mg/l) and IBA(5mg/l)showed moderate rate of callusing.

But when stem was cultured in BA (5mg/l) along with 2,4-D(5mg/l) in high concentration shows maximum rate of callus formation.

The maximum callus formation observed in 2,4-D containing medium.

Callus subculture

• Callus obtained from explants is subcultured on MS medium containgIAA(2mg/l) alongs with KIN(4mg/l).

• Auxin are known to promoterooting and sexual cases let as antogonostic to cytokinin. Recallusing was also observed.

SUMMARY AND CONCLUSION  Baliospermum montanum is a valuable medicinal plant Roots are used against purgative, anti-inflammatory, anodyne, digestive, diuretic. Pharmaceutical over exploitation and habitat destruction makes it endangered. Tissue culture approaches helps in the rapid production of the plant and there by plants can be

protected from extinction and also may be use full for the extraction of valuable product from the plant parts. 

Shoot tip cultures were used for plant regeneration. Maximum elongation of shoot tip was resulted in BA (5mg/l) and IBA (0.4mg/l) containing medium.

In nodal culture axillary bud proliferations was observed. Medium containing BA (4mg/l) and IAA (0.5mg/l) show maximum axillary bud elongation. IAA alone shows moderate callusing. Callus formation from the explant sources concentration and combination of the hormones for

callus induction explant such as stem were used. Stem shows maximum rate of callusing in medium containing BA (5mg/l) and alone with 2,4-D

(5mg/l)

Thank you……

midhun550@gmail.com