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Characterization of an algal phosphomannose isomerase gene and its application as a selectable marker for genetic manipulation of tomato

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Plant Diversity 2021年第1期43卷 63-70页

作者： Yuanyuan Lin Junchao Huang Key Laboratory of Economic Plants and Biotechnology Yunnan Key Laboratory for Wild Plant ResourcesKunming Institute of BotanyChinese Academy of SciencesKunming 650201People's Republic of China University of Chinese Academy of Sciences Beijing 100049People's Republic of China

Establishing a transgenic plant largely relies on a selectable marker gene that can confer antibiotic or herbicide resistance to plant *** existence of such selectable marker genes in genetically modified foods has long been *** cells generally exhibit too low an activity of phosphomannose isomerase(PMI)to grow with mannose as a sole carbon *** this study,we characterized PMI from the green microalga Chlorococcum *** assessed its feasibility as a selectable marker for plant *** ***(ChlPMI)was shown to be closely related to higher plants but more distant to bacterial *** of ChlPMI in tomato induced callus and shoot formation in media containing mannose(6 g/L)and had an average transformation rate of 3.9%.Based on this transformation system,a polycistronic gene cluster containing crtB,HpBHY,CrBKT and SlLCYB(BBBB)was co-expressed in a different tomato *** putative transformants were achieved with a transformation rate of 1.4%,which produced significant amounts of astaxanthin due to the expression of the BBBB *** together,these findings indicate that we have established an additional tool for plant biotechnology that may be suitable for genetically modifying foods safely.

关键词： Algae Tomato Transformation phosphomannose isomerase Astaxanthin

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Using the phosphomannose isomerase (PMI) Gene from Saccharomyces cerevisiae for Selection in Rice Transformation

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Journal of Integrative Agriculture 2012年第9期11卷 1391-1398页

作者： WANG Tao LIU Liang-yu TANG Yong-yan ZHANG Xiao-bo ZHANG Mei-dong ZHENG Yong-lian ZHANG Fang-dong National Key Laboratory of Crop Genetic Improvement Ministry of Science and Technology/Huazhong Agricultural University Wuhan 430070P. R. China

The phosphomannose isomerase （PMI） gene from Saccharomyces cerevisiae acted as selectable marker and mannose acted as selective agent for the production of transgenic plants of rice （Oryza sativa L.） via Agrobacterium-mediated transformation. The concentration of mannose during the selection was stepwise increased, 5 g L-1 mannose combined with 15 g L-1 sucrose and 500 mg L-1 cefotaxime was used in the initial selection stage, then the concentration of mannose was increased to 11 g L-1, the highest transformation rate was 20.0%. The integration of PMI gene was confirmed by PCR, and the result of RT-PCR assay proved that the intron of PMI gene can be excised correctly during RNA splicing. 13- Glucuronidase （GUS） activity analysis confirmed the expression of GUS gene. All those means the PMI gene from yeast can be used as a selectable marker in rice transformation.

关键词： phosphomannose isomerase selectable marker Saccharomyces cerevisiae transformation rice

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