Skaliter Oded, Ravid Jasmin, Shklarman Elena, Ketrarou Nadav, Shpayer Noam, Ben Ari Julius, Dvir Gony, Farhi Moran, Yue Yuling, Vainstein Alexander
The Robert H. Smith Faculty of Agriculture, Food and Environment, Institute of Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, Rehovot, Israel.
The Laboratory for Mass Spectrometry and Chromatography, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel.
Front Plant Sci. 2019 Nov 27;10:1561. doi: 10.3389/fpls.2019.01561. eCollection 2019.
Floral pigmentation is of major importance to the ornamental industry, which is constantly searching for cultivars with novel colors. Goldenrod () has monochromatic yellow carotenoid-containing flowers that cannot be modified using classical breeding approaches due to a limited gene pool. To generate Solidago with novel colors through metabolic engineering, we first developed a procedure for its regeneration and transformation. Applicability of different cytokinins for adventitious regeneration was examined in the commercial cv. Tara, with zeatin yielding higher efficiency than 6-benzylaminopurine or thidiazuron. A comparison of regeneration of commercial cvs. Tara, Golden Glory and Ivory Glory revealed Tara to be the most potent, with an efficiency of 86% (number of shoots per 100 leaf explants). Agrobacterium-based transformation efficiency was highest for cv. Golden Glory (5 independent transgenic shoots per 100 explants) based on kanamycin selection and the GUS reporter gene. In an attempt to promote anthocyanin biosynthesis, we generated transgenic Solidago expressing snapdragon () Rosea1 and Delila, as well as PRODUCTION OF ANTHOCYANIN PIGMENT 1 (PAP1) transcription factors. Transgenic cv. Golden Glory expressing cauliflower mosaic virus 35S-driven generated red flowers that accumulated delphinidin and its methylated derivatives, as compared to control yellow flowers in the GUS-expressing plants. The protocol described here allows efficient engineering of for novel coloration and improved agricultural traits.
花卉色素沉着对观赏产业至关重要，该产业一直在寻找具有新颖颜色的品种。一枝黄花（）具有含单一黄色类胡萝卜素的花朵，由于基因库有限，无法使用传统育种方法对其进行改良。为了通过代谢工程培育出具有新颖颜色的一枝黄花，我们首先开发了其再生和转化程序。在商业品种cv. Tara中检测了不同细胞分裂素对不定芽再生的适用性，玉米素的效率高于6-苄基腺嘌呤或噻苯隆。对商业品种cv. Tara、Golden Glory和Ivory Glory的再生进行比较，发现Tara最有效，效率为86%（每100个叶片外植体的芽数）。基于卡那霉素选择和GUS报告基因，基于农杆菌的转化效率在cv. Golden Glory中最高（每100个外植体有5个独立的转基因芽）。为了促进花青素生物合成，我们培育了表达金鱼草（）Rosea1和Delila以及花青素色素1（PAP1）转录因子的转基因一枝黄花。与表达GUS的植物中的对照黄花相比，表达花椰菜花叶病毒35S驱动的的转基因cv. Golden Glory产生了积累飞燕草素及其甲基化衍生物的红色花朵。这里描述的方案允许对进行高效工程改造以获得新颖的颜色和改善农业性状。
