LIU Xiaowei,LI Houhua,WANG Bingjie,et al.Construction of Recombinant Vector of AmAS 1 and Am4’CGT Genes and the Transient Expression in Malus Flowers[J].Northern Horticulture,2016,40(14):97-102.[doi:10.11937/bfyy.201614024]
AmAS1与Am4’CGT基因双价载体构建及在海棠花中的瞬时表达
- Title:
- Construction of Recombinant Vector of AmAS 1 and Am4’CGT Genes and the Transient Expression in Malus Flowers
- Keywords:
- crabapple; aurones; bivalent expression vector; transient expression
- 文献标志码:
- A
- 摘要:
- 以黄色金鱼草（Antirrhinum majus）、“雪球”海棠（Malus ‘Snow drift’）为试材，采用农杆菌介导法，将构建的金鱼草素合酶基因（aureusidin synthase,AmAS1）和查尔酮4’-O-葡萄糖苷转移酶基因（chalcone 4’-O-glucosyltransferase，Am4’CGT）双价表达载体在“雪球”活体花瓣上进行瞬时表达，并通过高效液相色谱分析和基因表达分析对瞬时表达效果进行鉴定。结果表明：成功构建双价表达载体CGT-AS-pCambia 1302,农杆菌侵染“雪球”花苞后，花苞呈现淡黄色，表明目的基因已在花瓣内转录并表达。高效液相色谱分析发现，侵染后的花苞出现新的吸收峰，进一步确定花瓣内有橙酮物质的合成。
- Abstract:
- Taking Antirrhinum majus and Malus ‘Snow drift’ as materials，using agrobacterium-mediated method，the recombinant vector containing Am4’CGT and AmAS1 was expressed in Malus flowers，the instantaneous expression effect was identified by HPLC and PCR analysis.The results showed that the recombinant vector CGT-AS-pCambia 1302 was built successfully，after infected by EHA105，the Malus flowers appeared to be light yellow，and it indicated that the target genes were transcribed and expressed in Malus flowers.A new absorption peak was found by HPLC analysis，the result indicated that there were aurones accumulated.
参考文献/References:
[1]李厚华，赵冰，李鹏民.中国海棠[M].北京:中国林业出版社，2014:1-13. [2]高立旦，童再康，曹件生，等.彩叶木本植物育种研究进展[J].浙江林业科技，2006，26(4)：77-82. [3]TANAKA Y，SASAKI N，OHMIYA A.Biosynthesis of plant pigments：anthocyanins，betalains and carotenoids[J].The Plant Journal，2008,54：733-749. [4]周琳，王雁，彭镇华.黄色花形成机制及基因工程研究进展[J].林业科学，2009，45(2)：111-119. [5]TANAKA Y，ONO E，NAKAMURA N，et al.Method for producing yellow flower by controlling flavonoid synthetic pathway:US 7750209 B2[P].U.S.Patent，International Flower Developments Proprietary Limited. [6]NAKAYAMA E.Enzymology of aurone biosynthesis[J].Journal of Bioscience and Bioengineering,2002,94(6)：487-491. [7]NAKAYAMA T，SATO T，FUKUI Y，et al.Specificity analysis and mechanism of aurone synthesis catalyzed by aureusidin synthase，a polyphenol oxidase homolog responsible for flower coloration[J].FEBS Letters，2001，499(1-2)：107-111. [8]ONO E，FUKUCHI-MIZUTANI M，NAKAMURA N，et al.Yellow flowers generated by expression of the aurone biosynthetic pathway[J].Proceedings of the National Academy of Sciences of the United States of America，2006，103(29)：11075-11080. [9]KAPILA J，RYCKE R D，MONTAGU M V，et al.An agrobacterium mediated transient gene expression system intact leaves[J].Plant Science，1997,122：101-108. [10]ZHANG Y Z，LI P M，CHENG L L.Developmental changes of carbohydrates，organic acids，amino acids，and phenolic compounds in ‘Honeycrisp’ apple flesh[J].Food Chemistry，2010,123：1013-1018. [11]宋建，刘仲齐.农杆菌介导的基因瞬时表达技术及其应用[J].天津农业科学，2008,14(1)：20-22. [12]邱礽，陶刚，李奇科，等.农杆菌渗入法介导的基因瞬时表达技术及应用[J].分子植物育种,2009，7(5)：1032-1039. [13]NAKAYAMA T，YONEKURA-SAKAKIBARA K，SATO T，et al.Aureusidin synthase：A polyphenol oxidase homolog responsible for flower coloration[J].Science，2000,290(5494)：1163-1166. [14]OREN S M，LEVI N A.Temperature effect on leaf pigmentation of Cotinus coggygri ‘Royal Purple’[J].Journal of Horticulture Science，1997,72：425-432.
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备注/Memo
第一作者简介：刘小微（1988-），女，河北廊坊人，硕士研究生，研究方向为植物分子生物学。E-mail：1030742965@qq.com.责任作者：李厚华（1973-），男，山东济宁人，博士，副教授，硕士生导师，现主要从事植物基因工程和类黄酮次生代谢等研究工作。E-mail:lihouhua73@163.com.基金项目：国家自然科学基金资助项目（K3050215130）；中央财政林业推广资助项目([2014]15)。