Increased tolerance to multiple abiotic stresses in plants is a key target for food security. OsSAP8 is a member of the stress-associated protein (SAP) gene family in rice and is characterized by the presence of two zinc finger domains, A20 and AN1. In this research, we aim to understand the regulatory mechanism of OsSAP8 in response to multiple stresses for rice survival. We used the Arabidopsis transformation system to understand the gene's function. Two Arabidopsis lines were used; OsSAP8 overexpression line and Arabidopsis atsap2 knockout loss-of-function mutation in comparison to wild type Columbia-0 (Col-0). The protein sequence of Arabidopsis atsap2 has been shown to be highly identical to the OsSAP8. An atsap2 knockout showed a sensitive phenotype to high concentration of NaCl and PEG with reduced germination percentage. OsSAP8 overexpression showed a tolerance phenotype to 150 mM NaCl and 20% PEG compared to atsap2. OsSAP8 also showed an insensitive germination phenotype to high concentration of ABA compared to Col-0 and atsap2. The insensitive phenotype of OsSAP8 on the high concentration of ABA (12 µM) and the sensitive phenotype of its orthologue atsap2 in Arabidopsis suggests that OsSAP8 may have a role in part through ABA signaling as a putative transcriptional regulator for ABA during stress response.
Transformasi Gen Proteolisis 6 (PRT6) Berperantarakan Agrobacterium tumefaciens ke dalam Kotiledon Tomato kultivar Micro Tom (Agrobacterium tumefaciens Mediated Transformation of the Proteolysis 6 (PRT6) Gene into Cotyledons of Tomato cv. Micro Tom) INTAN ELYA SUKA, NUR FARHANA ROSLAN, BEE LYNN CHEW, HOE HAN GOH, ZAMRI ZAINAL & NURULHIKMA MD ISA* ABSTRAK Gen Proteolisis 6 (PRT6) merupakan gen yang memainkan peranan penting dalam tapak jalan N-end rule dan berfungsi sebagai enzim E3 ligase. PRT6 berperanan dalam pengenalan protein sasaran bagi proses degradasi. Objektif utama kajian ini adalah untuk mentransformasi konstruk RNAi PRT6 ke dalam tomato berperantarakan Agrobacterium tumefaciens. Ini bertujuan untuk memahami peranan tapak jalan N-end rule semasa proses pemasakan buah. Beberapa faktor yang memberi kesan kepada transformasi seperti masa ko-penanaman dan juga kepekatan antibiotik yang digunakan telah dioptimumkan. Keputusan kajian menunjukkan pengeraman kotiledon selama 48 jam pada medium ko-penanaman dapat meningkatkan penghasilan kalus sebanyak 61% manakala penggunaan 500 mg/L antibiotik karbenisilin dalam medium regenerasi pucuk dapat mengurangkan kontaminasi A. tumefaciens sehingga 5.2%. Selain itu, strain A. tumefaciens C58 merupakan strain A. tumefaciens yang paling sesuai digunakan sebagai perantara dalam kajian ini. Tindak balas berantai polimerase (PCR) telah dijalankan pada pucuk yang terhasil untuk mengesahkan integrasi fragmen PRT6 ke dalam genom tomato. Berdasarkan analisis PCR, kesemua tujuh pucuk putatif transgenik adalah merupakan transforman positif.
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