Improvement of freezing tolerance of lettuce by modification of low-temperature-inducible genes

Research Project

Project/Area Number	15K07456
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Food science
Research Institution	Kyushu University
Principal Investigator	Honjoh Ken-ichi 九州大学, 農学研究院, 准教授 (00264101)
Co-Investigator(Kenkyū-buntansha)	宮本敬久九州大学, 農学研究院, 教授 (70190816)
Project Period (FY)	2015-04-01 – 2018-03-31
Project Status	Completed (Fiscal Year 2017)
Budget Amount *help	¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000) Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywords	凍結耐性 / レタス / GolS / VPE
Outline of Final Research Achievements	cDNA cloning of GolS (LsGolS1, LsGolS2) gene and VPE (LsVPE) gene from lettuce was isolated to determine the nucleotide sequences. The LsGolS2 gene was expressed in Escherichia coli as a fusion protein with TF-His-tag, purified by enzyme treatment and so on, to obtain a protein of about 39 kDa. The substance serving as a substrate was added to this protein and the reaction product was examined by HPLC. As a result, formation of galactinol was found meaning that the activity of this protein was confirmed. Introduction of GolS gene into higher plant expression vector pRI101AN and introduction into lettuce using Agrobacterium. As a result of screening with PCR, transgenic lettuce was acquired.