Dissertation
Dissertation > Agricultural Sciences > Crop > Tuber Crops > Potatoes ( potatoes )

Research of Chlorogenic Acid-improved Transgenic Potato with AtMYB12

Author DaiLiLi
Tutor DingXinHua
School Shandong Agricultural University
Course Plant Pathology
Keywords Potato ’lu yin1hao’ Transgenic Chlorogenic-acid Antioxidation Antibacterium
CLC S532
Type Master's thesis
Year 2013
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The cultivated potato(Solanum tuberosum)belongs to the Solanaceae plant family,which is the most important vegetable and the fourth most important food crop in theworld.‘lu yin1hao’is a new potato cultivar which was introduced by the Shandong Academyof Agricultural Sciences from Holland. This new variety had been widely planted inShandong Province for the advantage of early maturity, high yield and quality. Chlorogenicacid is a natural polyphenolic compound which has antioxidative, anti-bacterial andcardiovascular protection functions, but only small amounts can be detected on widelyplanting potato varieties, and there are significant differences among different varieties.Potatoe is an autotetraploid genome plant, the method of cross-breeding of improvedvarieties are more difficult. One of goals of the genetics and breeding is to improve nutritionalquality. Potato can be vegetatively propagated by tubers and transgenic traits passed on tofuture generations without purification or anther culture breeding,so utilizing geneticengineering techniques to cultivate new potato varieties is a immense advantage. Studies haveshown that transcription factors can improve the content of secondary metabolites in plants,with the deepening study of molecular biology, potato genetic transformation system hadbeen matured. Utilizing HPLC and mass spectrometry method, We detected the content ofchlorogenic acid in potato ‘Desiree’ tubes carrying AtMYB12gene. The potato carriedpX6-patatin::AtMYB12, the content of chlorogenic acid is rang from1.095to4.846mg g-1.In this study, the AtMYB12gene cloned from Arabidopsis was transferred into potato(Solanum tuberosum L.) cultivars ‘lu yin1hao’ via agro bacterium mediated stem segmenttransformation. In total,25, positive transgenic plant individuals were obtained with32%oftransformation efficiency.Utilizing HPLC method, we detected the content of chlorogenic acid in potato carriedpX6-patatin:: AtMYB12, the content of chlorogenic acid is range from1.613to3.693mg/gDW. Content was increased nearly20-fold compared to non-transgenic potato. Nowtransgenic security issues controversial, so we based chlorogenic acid content analysis and thestability of morphological traits,2lines selected transgenic combination were treated withβ-estradiol, and were tested by PCR to obtain NPTII marker-free plants. We also analysis thecontent of chlorogenic acid, anti-oxidant and anti-bacterial activity of transgenic potato. Theresult indicated that the percentage of marker-free in transgenic combinations is4%.We detected the marker-free transgenic potato which carried AtMYB12gene, content ofCGA is range from1.766to3.748mg/g, anti-oxidant capacity increased by two times, the extract of transgenic tuber inhibition E. coli is very obvious compared with the blank.Byusing qRT-PCR to detect the expression level of genes DFR,FLS, HQT. The expression levelof three genes is increased. FLS gene expression levels up to23times compared withnon-transgenic transgenic potato; DFR gene expression amount of the highest non-transfected genes DFR gene expression amount of17times; HQT gene expression amount of60times multiples of transgenic potato HQT gene expression amount to improve,The resultindicated the selected gene was transferred to the complete potato varieties ‘lu yin1hao’.In summary, we have obtained new chlorogenic acid-rich and high anti-oxidant andanti-bacterial varieties by transgenic strategy, which is expected tube further screened in thefield. This study provides theoretical basis to obtained chlorogenic acid-rich potato varietiesand clear synthesis pathway of chlorogenic acid in potato and communication of the basis forsafety assessment, and to address the key problem in production.

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