Molecular genetic study of genes related to sucrose synthesis in cassava

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Oct 23, 2013 (3 years and 7 months ago)

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Molecular genetic study of genes related to sucrose synthesis in cassava


Kessarin Tungngoen, Supatcharee Netrphan, Malinee Suksangpanomrung and Jarunya Narangajavana

Department of Biotechnology, Faculty of Science, Mahidol University, Rama6 Road, Bangkok,

10400, Thailand and
National Center for Genetic Engineering and Biotechnology, 113 Paholyothin Rd., Klong 1, Klong Luang,
Patumthani 12120, Thailand



Sucrose is an important molecule, which fulfils many roles in starch biosynthesis, such as
a mediator fo
r transporting sugar between sink and source tissues as well as a supplier for a
storage reserve in source tissues. The enzymes that are known to involve in the synthesis of
sucrose include sucrose
-
phosphate synthase (SPS; EC 2.3.1.14), sucrose
-
phosphatase

(SPP; EC
3.1.3.24) and 14
-
3
-
3 protein. To elucidate the roles of these enzymes in sucrose transport
machinery in cassava (Manihot esculenta Crantz L.), the expression profile of SPS, SPP and 14
-
3
-
3 protein in various tissues and developmental stages, and
environmental conditions (e.g. light
and dark) will be investigated in this research. Using Reverse Transcription (RT)
-
PCR strategy,
the primers specific to SPS, SPP and 14
-
3
-
3 genes were synthesized based on the consensus
sequences observed in various pla
nt species. A partial cDNA fragment of SPS (623
-
bp) was
cloned from tuberous root tissues of cassava while partial sequences of SPP (460
-
bp) and 14
-
3
-
3
(373
-
bp) cDNA were cloned from leaves. Nucleotides sequence comparison revealed that the
cassava SPS cDN
A fragment shared 82% similarity to SPS mRNA from sweet potato while the
cassava SPP and 14
-
3
-
3 cDNA fragments shared 82% and 77% similarity with the SPP and 14
-
3
-
3 mRNA from Arabidopsis thaliana and tomato, respectively. These amplified cDNA fragments
wi
ll be used as probes in Northern blot analysis to determine whether the expression of cassava
SPS, SPP and 14
-
3
-
3 are regulated in tissue
-
, developmental
-

and light
-
dependent manners.


Acknowledgement

This research was supported by the Ministry of Universi
ty Affairs, under the Science and
Technology Higher Education Development Project in Agricultural Biotechnology, and by the
National Center for Genetic Engineering and Biotechnology (BIOTEC) No. BT
-
B
-
09
-
PG
-
BC
-
4605 and No. BT
-
B
-
09
-
PG
-
BC
-
4606.