CRISPR/Cas9-mediated mutagenesis of the dihydroflavonol-4-reductase-B (DFR-B) locus in the Japanese morning glory Ipomoea (Pharbitis) nil.
CRISPR/Cas9-mediated mutagenesis of the dihydroflavonol-4-reductase-B (DFR-B) locus in the Japanese morning glory Ipomoea (Pharbitis) nil.
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二氢类黄酮-4-还原酶-B（DFR-B）基因座的CRISPR/CAS9介导的诱变，日本早晨荣耀ipomoea（Pharbitis）nil。
DOI：
10.1038/s41598-017-10715-1
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发表时间：
2017-08-30
影响因子：
4.6
通讯作者：
Ono M
中科院分区：
文献类型：
--
作者：
Watanabe K;Kobayashi A;Endo M;Sage-Ono K;Toki S;Ono M
CRISPR/Cas9技术是一种在包括植物在内的许多生物中进行靶向突变的通用工具。然而，这项技术还没有应用于日本牵牛花(Ipomoea[Pharbitis]nil)，这是日本国家生物资源计划选择的一种传统园林植物。我们选择了编码花青素生物合成酶的近红花落叶松二氢黄酮醇-4-还原酶B(DFR-B)作为目的基因，通过根瘤菌[农杆菌]介导的转化，观察了在植物组织培养的早期阶段茎颜色的变化。在32株转基因植株中，有24株(75%)的花青素含量较低，在DFR-B的Cas9裂解位点有双等位基因突变，表现为一个碱基插入或两个以上碱基缺失。因此，这些结果表明，CRISPR/CAS9技术能够在该模式园艺植物中探索基因功能。据我们所知，这是第一个利用CRISPR/CAS9技术研究高等植物花色变化的报告。
CRISPR/Cas9 technology is a versatile tool for targeted mutagenesis in many organisms, including plants. However, this technique has not been applied to the Japanese morning glory (Ipomoea [Pharbitis] nil), a traditional garden plant chosen for the National BioResource Project in Japan. We selected dihydroflavonol-4-reductase-B (DFR-B) of I. nil, encoding an anthocyanin biosynthesis enzyme, as the target gene, and changes in the stem colour were observed during the early stages of plant tissue culture by Rhizobium [Agrobacterium]-mediated transformation. Twenty-four of the 32 (75%) transgenic plants bore anthocyanin-less white flowers with bi-allelic mutations at the Cas9 cleavage site in DFR-B, exhibiting a single base insertion or deletions of more than two bases. Thus, these results demonstrate that CRISPR/Cas9 technology enables the exploration of gene functions in this model horticultural plant. To our knowledge, this report is the first concerning flower colour changes in higher plants using CRISPR/Cas9 technology.
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影响因子：
46.9
作者：
Doench JG;Fusi N;Sullender M;Hegde M;Vaimberg EW;Donovan KF;Smith I;Tothova Z;Wilen C;Orchard R;Virgin HW;Listgarten J;Root DE
通讯作者：
Root DE
影响因子：
11.6
作者：
Hayama, Ryosuke;Agashe, Bhavna;Coupland, George
通讯作者：
Coupland, George
影响因子：
16.6
作者：
Hoshino, Atsushi;Jayakumar, Vasanthan;Nitasaka, Eiji;Toyoda, Atsushi;Noguchi, Hideki;Itoh, Takehiko;Shin-, Tadasu, I;Minakuchi, Yohei;Koda, Yuki;Nagano, Atsushi J.;Yasugi, Masaki;Honjo, Mie N.;Kudoh, Hiroshi;Seki, Motoaki;Kamiya, Asako;Shiraki, Toshiyuki;Carninci, Piero;Asamizu, Erika;Nishide, Hiroyo;Tanaka, Sachiko;Park, Kyeung-Il;Morita, Yasumasa;Yokoyama, Kohei;Uchiyama, Ikuo;Tanaka, Yoshikazu;Tabata, Satoshi;Shinozaki, Kazuo;Hayashizaki, Yoshihide;Kohara, Yuji;Suzuki, Yutaka;Sugano, Sumio;Fujiyama, Asao;Iida, Shigeru;Sakakibara, Yasubumi
通讯作者：
Sakakibara, Yasubumi
影响因子：
64.8
作者：
Fukada-Tanaka, S;Inagaki, Y;Iida, S
通讯作者：
Iida, S
DOI：
10.1073/pnas.1400822111
发表时间：
2014-03-25
影响因子：
11.1
作者：
Feng, Zhengyan;Mao, Yanfei;Zhu, Jian-Kang
通讯作者：
Zhu, Jian-Kang