MR. WITTAYA PROMME - Rubber Boardclinic.rubberboard.org.in/PDF/ircpdf/BT-07 WITTAYA.pdf · MR. WITTAYA PROMME In Vitro Micropropagation of ... Calopogonium mucunoides Calopogonium

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MR. WITTAYA PROMME

In Vitro Micropropagation ofCa l opogon i um cae ru l eumFor Soil Cover Crop underR u b b e r P l a n t a t i o n

Chacheongsao Rubber Research CenterRubber Research Institute of Thailand,Department of Agriculture

Their beneficial effects

INTRODUCTIONA mixture of legumes species is planted as a soilcover in the intercrop.

1 •Nitrogen fixation

2 •Nutrient return 

3 • Soil erosion control

4 •Nutritive value

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The soil covers in rubber plantation effect tosoil fertility, integrate with compatible grassesand the growth of rubber tree.

The creeping legumes widely used ascover crops under rubber plantation

Pueraria phaseoloides

Centrosema pubescens

Calopogonium mucunoides

Calopogonium caeruleum

Calopogonium caeruleum is new

one of the cover crop important in rubberplantation !!

- It is a tropical pastures legume under plantation.

-It is higher shad tolerance than the both of cover crops, Calopogonium mucunoides and Centrosema pubescens(medium shade tolerance)

-It is one of a species suitable for soil cover under plantation

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The main problems ofCalopogonium caeruleum : 

Low seed setHigh cost: 10-17 USD/kg seed

We found no information in theliterature pertaining to the in vitroculture of Calopogonium caeruleum.However, has been reported in vitroculture from several other legume species.

In this study to establish a highfrequency plant regeneration system fromCalopogonium caeruleum for more produceyoung plants for soil cover crop underrubber plantation.

Study Objective….

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EXPLANTS INITIATION AND ESTABLISHMENT

OF ASEPTIC CULTURE

The preparation of seed coat breakand contamination free explants.

Preparation of seed coat break

Seed coat break usingconcentrate sulfuricacid for 5-10 min,treatment for maximumgermination.

Contamination free explants

-Washing with distilled water for 3-5times.

-Washing with 70 % alcohol, 10 min anddouble sterilization with 20 % (v/v)Clorox, 10 min and 10 % (v/v) Clorox, 5min.

-Washing in sterile water 3-4 time

-Left to air dry.

The seed coat broke seedswere surface sterilized by …..

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Seed were culturedon medium…..

Sterilized seeds were placed onMurashige and Skoog medium(MS) supplemented with 0.5 -2.0mg/l BA

Germinate at dark chamber for3-days and then in light for 4-days, maximum germination 85-100 %

MULTIPLE SHOOT INDUCTION

The maximum response notified incotyledonary node explants wasevaluated as the mean number of shootsof 7.

Cotyledonary node were cultured on MSmedium supplemented with 1.0-2.0 mg/lBA and 0.25- 0.50 mg/l NAA for multipleshoot induction.

Cotyledonary node were excised fromthe surface sterilized, in vitro grown, 7-d old seedlings.

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ROOTING OF THE IN VITRO SHOOTS

MS medium + 1 mg/l IAA was found maximumnumber of rooting (6 roots/shoot)

MS medium 1 mg/l IBA or IAA was

found to be more effective in the production oflong, slender and healthy roots

MS hormone free medium was found maximumof roots length is (4.0 cm), whereas MSmedium supplemented with IBA, IAA and NAAwas found 2.9, 1.6 and 0.4 cm.

MS hormone free medium and MS + 1 mg/l IBAor IAA was found to be more effective in theproduce maximum rooting (100 %)

The survival rate of the plants :The field humidity conditions controlwas recorded as 80 %, whereas55 % survival rate without control.

HARDENING OF YOUNG PLANTLETS

:The field humidity conditionscontrol and:The field humidity conditionswithout control.

Rooted plantlets were initiallyhardened under

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THANK YOU