The development of molecular biology techniques nowadays has enabled to engineer drought tolerant sugarcane by genetic engineering to accelerate the breeding program. Dehydrin (DHN) is known to have an important role in plant response and adaptation to abiotic stresses (drought, high salinity, cold, heat, etc.). While plant tissues are subjected to drought stress (dehydration), DHN protein is accumulated to high content throughout all vegetative or generative tissues. The research aimed to isolate and characterize the DHN promoter from sugarcane that can be used as transformation material in generating drought tolerant sugarcane. Specific primers for DHN promoter amplification were designed and DHN promoter region was successfully isolated by PCR cloning method. Two putative promoter sequences were identified namely Pr-1DHNSo and Pr-2DHNSo. In silicoanalyses were carried out and cis-regulatory elements motifs that play a role in adaptation on abiotic stress as well as biotic stress including ABRE, MBS, CGTCA-motif, TGACG-motif, GARE-motif, P-box TCA-element and Box-W1 were identified. The promoter Pr-1DHNSo was then cloned into pBI121 expression vector by Overlap Extention PCR (OE-PCR) for further characterization. Functional test of the promoter construct pBI- Pr-1DHNSo was conducted through Agrobacterium transformation into sugarcane calli. GUS assay and PCR analysis showed that the DHN promoter was transformed and expressed in the sugarcane calli.
Extreme climate change requires rice varieties adaptable to drought condition. Adaptation will play an important role in ensuring the sustainability of food security. This research aimed to improve drought tolerance of Inpari 30 and Situ Bagendit varieties through Spike-Stalk Injection Method (SIM). DNAs from several plant species, such as rice (cv. Cabacu), grasses (Echinochloa crusgalli/E. colona, elephant grass/Pennisetum purpureum, Bothriochloa pertusa, Cenchrus echinatus, Sorghum nitidum, Ischamemum timorensis and Guinea grass), sugarcane, wild rice (Oryza nivara), maize and sorghum were injected to tillers of cultivar Inpari 30 and Situ Bagendit. The first set of M1 Situ Bagendit-SIM and M1 Inpari 30-SIM seeds were treated with 20% of PEG 8000 solution for 10 days. Some of the well germinated seeds were planted in pots and maintained untill harvest. The selected M2 Situ Bagendit-SIM and M2 Inpari 30-SIM and the second set of M1 Situ Bagendit-SIM and M1 Inpari 30-SIM seeds were planted at Muara Experimental Station, Bogor (West Java). The result of PEG 8000 assay showed that M1 Situ Bagendit-SIM-elephant grass, sugarcane and O. nivara had significantly longer radicle length and higher fresh weight compared to Situ Bagendit, while M1 Inpari 30-SIM-sugarcane, sorghum and elephant grass had significantly longer radicle and plumule length, and higher radicle weight compared to Inpari 30. Field trial showed that the mutant lines of Situ Bagendit-SIM performed better than those of Inpari 30-SIM. The grain weight of M1 Situ Bagendit-SIM-Cabacu, jajagoan grass, maize, sugarcane, O. nivara and B. pertusa, and M2 Situ Bagendit-SIM-O. nivara and elephant grass were higher than that of Situ Bagendit. The grain weight of M1 Inpari 30-SIM-jajagoan grass was also higher than that of Inpari 30. Therefore, SIM could be an alternative way to develop genetic variation of rice plant.
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