Oil palm MSP2 promoter isolation, in silico analysis and functional characterisation

Nurniwalis Abdul Wahab and Zubaidah Ramli and Mohamad Arif Abd Manaf and Ahmad Parveez Ghulam Kadir (2023) Oil palm MSP2 promoter isolation, in silico analysis and functional characterisation. Journal of Oil Palm Research (Malaysia), 35 (3). pp. 491-503. ISSN 2811-4701

Arabidopsis was used as a stable transformation system to characterise the function of the oil palm mesocarpspecific promoter, MSP2. Five MSP2 promoter fragments ranging from 1558 bp to 3044 bp in size were successfully isolated. In silico sequence analysis showed various putative plant regulatory elements in the different MSP2 promoter regions. Two vector constructs containing MSP2-GLC and MSP2-GLG promoter fragments were attached to beta-glucuronidase (GUS) reporter gene and transformed into Arabidopsis thaliana. Histochemical GUS staining of the transgenic Arabidopsis showed that both promoter fragments were expressed in the flower, especially in petal and stigma, silique for MSP2-GLC and in anther and stamen for MSP2-GLG. The QELEMENTZMZM13 motif present only in the MSP2-GLG fragment is likely vital for the anther-specific expression. In MSP2-GLC transgenic Arabidopsis, GUS expression was enhanced under cold conditions, unlike MSP2-GLG transgenic Arabidopsis. A low-temperature response (LTR) motif present in MSP2-GLC may be important to enhance and drive the expression of a transgene under cold conditions. The deleted region in MSP2-GLG fragment caused the removal of the LTR motif, which most likely indicates that the size of the promoter is not necessarily important to drive gene expression, but the availability of a specific motif is key to determine its strength and specificity.

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Abstract

Arabidopsis was used as a stable transformation system to characterise the function of the oil palm mesocarpspecific promoter, MSP2. Five MSP2 promoter fragments ranging from 1558 bp to 3044 bp in size were successfully isolated. In silico sequence analysis showed various putative plant regulatory elements in the different MSP2 promoter regions. Two vector constructs containing MSP2-GLC and MSP2-GLG promoter fragments were attached to beta-glucuronidase (GUS) reporter gene and transformed into Arabidopsis thaliana. Histochemical GUS staining of the transgenic Arabidopsis showed that both promoter fragments were expressed in the flower, especially in petal and stigma, silique for MSP2-GLC and in anther and stamen for MSP2-GLG. The QELEMENTZMZM13 motif present only in the MSP2-GLG fragment is likely vital for the anther-specific expression. In MSP2-GLC transgenic Arabidopsis, GUS expression was enhanced under cold conditions, unlike MSP2-GLG transgenic Arabidopsis. A low-temperature response (LTR) motif present in MSP2-GLC may be important to enhance and drive the expression of a transgene under cold conditions. The deleted region in MSP2-GLG fragment caused the removal of the LTR motif, which most likely indicates that the size of the promoter is not necessarily important to drive gene expression, but the availability of a specific motif is key to determine its strength and specificity.

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Item Type:	Article
AGROVOC Term:	palm oils
AGROVOC Term:	mesocarp
AGROVOC Term:	plant biotechnology
AGROVOC Term:	Arabidopsis thaliana
AGROVOC Term:	research
AGROVOC Term:	staining
AGROVOC Term:	gene expression
AGROVOC Term:	genetic engineering
Geographical Term:	Malaysia
Uncontrolled Keywords:	Arabidopsis, Elaeis guineensis, heterologous system, mesocarp-specific promoter, model plant
Depositing User:	Nor Hasnita Abdul Samat
Date Deposited:	29 Apr 2025 01:10
Last Modified:	29 Apr 2025 01:10
URI:	http://webagris.upm.edu.my/id/eprint/1757

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