Cryopreservation of hairy root culture of Amrithapala, Decalepis arayalpathra (Joseph & Chandras) Venter
By: Dhanya C S.
Contributor(s): C G Sudha (Guide).
Material type:
BookPublisher: Vellayani Department of Plant Biotechnology, College of Agriculture 2014Description: 85p.Subject(s): Plant BiotechnologyDDC classification: 660.6 Online resources: Click here to access online Dissertation note: MSc(INT) Abstract: The present study, “Cryopreservation of hairy root culture of Decalepis
arayalpathra (Joseph & Chandras.) Venter” was carried out at the Biotechnology and
Bioinformatics Division, Jawaharlal Nehru Tropical Botanic Garden and Research
Institute, Palode during 2013-2014. The objective of the study is to find an efficient
cryopreservation protocol for hairy root culture of Decalepis arayalpathra and a
comparative analysis of secondary metabolite profile. Hairy roots of Decalepis
arayalpathra, a critically endangered medicinal plant used by Kani tribes as an effective
remedy for peptic ulcer and cancer like afflictions were attempted to cryopreserve using
encapsulation-dehydration and vitrification methods. A comparative analysis of
secondary metabolite profile of cryopreserved and normal hairy roots was also made
to examine practical utility of cryopreserved hairy roots. Encapsulation-dehydration
method was not good for preserving hairy roots of D. arayalpathra in liquid nitrogen.
The roots were highly sensitive to sucrose in pre-culture solution at least to get
dehydration tolerance. Encapsulated root tips pre-cultured with 0.5 M sucrose did not
survive even after one hour dehydration. Inclusion of DMSO in the pre-culture
solutions improved dehydration tolerance of encapsulated root tips but did not
support LN tolerance. Hairy root tips (2-3 mm) pre-cultured on half strength
Murashige and Skoog (MS) solid medium with 0.1 mg/l 2, 4-D for two days followed
by exposure to PVS2 for 45 min gave 50-100 percent recovery and regeneration after
cryopreservation. Roots regenerated from cryopreserved root tips grew like normal
hairy roots showing similar secondary metabolite profile as revealed by thin layer
chromatography. HPLC profiling of MBALD compound in cryopreserved root during
first subculture passage was less compared to that of the normal hairy root culture.
The study revealed cryopreservation by vitrification as a useful method to preserve
hairy root of D. arayalpathra.
| Item type | Current location | Collection | Call number | Status | Date due | Barcode |
|---|---|---|---|---|---|---|
Theses
|
KAU Central Library, Thrissur Theses | Reference Book | 660.6 DHA/CR (Browse shelf) | Not For Loan | 173438 |
MSc(INT)
The present study, “Cryopreservation of hairy root culture of Decalepis
arayalpathra (Joseph & Chandras.) Venter” was carried out at the Biotechnology and
Bioinformatics Division, Jawaharlal Nehru Tropical Botanic Garden and Research
Institute, Palode during 2013-2014. The objective of the study is to find an efficient
cryopreservation protocol for hairy root culture of Decalepis arayalpathra and a
comparative analysis of secondary metabolite profile. Hairy roots of Decalepis
arayalpathra, a critically endangered medicinal plant used by Kani tribes as an effective
remedy for peptic ulcer and cancer like afflictions were attempted to cryopreserve using
encapsulation-dehydration and vitrification methods. A comparative analysis of
secondary metabolite profile of cryopreserved and normal hairy roots was also made
to examine practical utility of cryopreserved hairy roots. Encapsulation-dehydration
method was not good for preserving hairy roots of D. arayalpathra in liquid nitrogen.
The roots were highly sensitive to sucrose in pre-culture solution at least to get
dehydration tolerance. Encapsulated root tips pre-cultured with 0.5 M sucrose did not
survive even after one hour dehydration. Inclusion of DMSO in the pre-culture
solutions improved dehydration tolerance of encapsulated root tips but did not
support LN tolerance. Hairy root tips (2-3 mm) pre-cultured on half strength
Murashige and Skoog (MS) solid medium with 0.1 mg/l 2, 4-D for two days followed
by exposure to PVS2 for 45 min gave 50-100 percent recovery and regeneration after
cryopreservation. Roots regenerated from cryopreserved root tips grew like normal
hairy roots showing similar secondary metabolite profile as revealed by thin layer
chromatography. HPLC profiling of MBALD compound in cryopreserved root during
first subculture passage was less compared to that of the normal hairy root culture.
The study revealed cryopreservation by vitrification as a useful method to preserve
hairy root of D. arayalpathra.
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