IN INOCULANTS Nodulaid - 17th International Nitrogen Fixation ...
IN INOCULANTS Nodulaid - 17th International Nitrogen Fixation ... IN INOCULANTS Nodulaid - 17th International Nitrogen Fixation ...
17 th International Congress on Nitrogen Fixation Fremantle, Western Australia 27 November – 1 December 2011 Title The role of 1-aminocyclopropane-1-carboxylate (ACC) deaminase enzyme on leguminous nodule senescence Author Sudarat Sripakdi 1 , Panlada Tittabutr 1 , Nantakorn Boonkerd 1 & Neung Teaumroong 1 Poster Board Number 23 1 School of Biotechnology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand. The objective of this study was to determine the role of 1-aminocyclopropane-1-carboxylate (ACC) deaminase of Ensifer sp. (Sinorhizobium sp.) strain BL3 on leguminous nodule senescence. The acdRS genes encoding ACC deaminase were cloned from BL3 into multiple copy plasmids (pRK404A), and transferred to wild type. The BL3 transconjugant containing of multiple copy number of acdRS (BL3 + ) greatly enhanced ACC deaminase activity compared to BL3 (1.634 and 0.447 μmol of �-ketobutyrate mg -1 protein h -1 , respectively). ACC deaminase activity of BL3 was also higher than mungbean nodulating strain as Bradyrhizobium sp. PRC008 (0.126 μmol of �-ketobutyrate mg -1 protein h -1 ). Moreover, BL3 + retarded nodule senescence (60.25%) greater than BL3 (39.75%) after 5 weeks planting. In the meantime, the kanamycin resistance gene (Km r ) were inserted into the acdRS fragment and transferred into BL3 in order to construct BL3 mutant, which was interrupted at acdR and acdS (BL3 - ). All strains were inoculated to the host plant, Vigna radiata cultivar SUT1. Inoculation of BL3, and BL3 + were not significantly effected on plant growth and root nodule number whilst BL3 - showed significantly lower than both strains. Whereas flowering period on BL3 + was longer than wild type and mutant strains. 138 2011
17 th International Congress on Nitrogen Fixation Fremantle, Western Australia 27 November – 1 December 2011 Title Gamma irradiation and autoclave sterilization of peat and compost as the carrier for rhizobial inoculant production Author Panlada Tittabutr 1 , Kamonluck Teamthisong 2 , Neung Teaumroong 1 & Poster Board Number 24 Nantakorn Boonkerd 1 1 School of Biotechnology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand 2 The Center for Scientific and Technological Equipment, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand Although several alternative carriers have been investigated and successfully used instead of peat, the main problem found in rhizobial inoculant production is contamination by other bacteria or molds that cause reduction of rhizobial cell number. This study aimed to elucidates the efficient sterilization process of two carriers, peat and compost before using as rhizobial inoculant. Peat and compost could be efficiently sterilized by irradiation. The carrier with 10% moisture content could be sterilized by irradiation at 10 kGy, while carrier with 30% moisture content must be sterilized by irradiation at 25 kGy. Penetration of irradiation through polypropylene (PP) bag tend to be better than polyethylene (PE) bag, since lower dose of irradiation was needed for carrier sterilization. However, PE bag appear to be more durable than PP bag after gamma irradiation at high doses. Nevertheless, contaminants could be detected in irradiated carrier after storage at room temperature for two months. Thus, autoclaving with tyndallization approach was applied in order to minimize the contaminant microorganisms in the carrier. Carriers with 10% moisture were autoclaved two times in a row at 121ºC for 60 min, with the waiting period during each time after autoclaving for 18 h could eliminate the fungal contaminants, while bacterial contaminants still remained about 10 2 cfu/g carrier. The number of Bradyrhizobium sp. PRC008 was in the range of 10 8 -10 9 cfu/g in both irradiated and autoclaved peat after 6 months storage. However, the numbers of bradyrhizobial cell were reduced in compost sterilized by both methods after one month storage. These results indicated that carrier material had an important influence on inoculant quality, while sterilization processes using gamma irradiation and autoclaving with tyndallization approach could be used for efficient rhizobial inoculant production with peat based carrier. 139 2011
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17 th <strong>International</strong> Congress on <strong>Nitrogen</strong> <strong>Fixation</strong><br />
Fremantle, Western Australia<br />
27 November – 1 December 2011<br />
Title The role of 1-aminocyclopropane-1-carboxylate (ACC) deaminase enzyme on leguminous<br />
nodule senescence<br />
Author Sudarat Sripakdi 1 , Panlada Tittabutr 1 , Nantakorn Boonkerd 1 & Neung Teaumroong 1<br />
Poster Board Number 23<br />
1 School of Biotechnology, Suranaree University of Technology, Nakhon Ratchasima<br />
30000, Thailand.<br />
The objective of this study was to determine the role of 1-aminocyclopropane-1-carboxylate (ACC) deaminase of<br />
Ensifer sp. (Sinorhizobium sp.) strain BL3 on leguminous nodule senescence. The acdRS genes encoding ACC<br />
deaminase were cloned from BL3 into multiple copy plasmids (pRK404A), and transferred to wild type. The BL3<br />
transconjugant containing of multiple copy number of acdRS (BL3 + ) greatly enhanced ACC deaminase activity<br />
compared to BL3 (1.634 and 0.447 μmol of �-ketobutyrate mg -1 protein h -1 , respectively). ACC deaminase<br />
activity of BL3 was also higher than mungbean nodulating strain as Bradyrhizobium sp. PRC008 (0.126 μmol of<br />
�-ketobutyrate mg -1 protein h -1 ). Moreover, BL3 + retarded nodule senescence (60.25%) greater than BL3<br />
(39.75%) after 5 weeks planting. In the meantime, the kanamycin resistance gene (Km r ) were inserted into the<br />
acdRS fragment and transferred into BL3 in order to construct BL3 mutant, which was interrupted at acdR and<br />
acdS (BL3 - ). All strains were inoculated to the host plant, Vigna radiata cultivar SUT1. Inoculation of BL3, and<br />
BL3 + were not significantly effected on plant growth and root nodule number whilst BL3 - showed significantly<br />
lower than both strains. Whereas flowering period on BL3 + was longer than wild type and mutant strains.<br />
138<br />
2011
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