乙烯利和1-MCP对菠萝蜜果实中AheAAT和AheERF表达的影响  

Effects of Ethephon and 1-MCP on the Expression of AheAAT Gene and AheERF Transcription Factors in Jackfruit Fruit

作　　者：任雪岩 刘光财 李国鹏[1] 叶春海[1] 丰锋[1] 王俊宁[1] REN XueYan;LIU GuangCai;LI GuoPeng;YE ChunHai;FENG Feng;WANG JunNing(Agricultural College of Guangdong Ocean University,Zhanjiang 524088,Guangdong)

机构地区：[1]广东海洋大学农学院

出　　处：《中国农业科学》2019年第21期3890-3902,共13页Scientia Agricultura Sinica

基　　金：国家自然科学基金青年基金(31401928);广东省自然科学基金博士启动项目(S2013040011544);广东海洋大学引进人才科研启动项目(1312130);广东海洋大学硕士学位论文培育项目(201841);广东海洋大学配套经费项目(C15486,C15491);广东海洋大学大学生创新项目(CXXL2014068)

摘　　要：【目的】研究乙烯(ETH)和1-甲基环丙烯(1-MCP)处理对菠萝蜜果实成熟过程中醇酰基转移酶(AAT)活性、AheAAT和AheERF1/2转录因子表达的影响,为进一步认识菠萝蜜果实AATs在酯类物质合成中的作用及其调控提供理论依据。【方法】以‘海大2号’菠萝蜜果实为试材,果实盛花期选择具有代表性、花期一致的果实挂牌,花后约150 d采收,选择大小和成熟度相同,且无病虫伤的果实,分成3组进行处理。第一组采用1000 mg·L-1 ETH溶液浸泡2—3 min;第二组采用0.5 mg·L-11-MCP熏蒸处理15 h;第三组作为对照组。在果实成熟过程中定期取样,每次取样3—5个果,测定AAT酶活性、AheAAT和AheERF表达等指标。【结果】通过测定AAT酶活性发现,ETH处理促进了果实的成熟衰老,提前了AAT活性高峰出现的时间,但是降低了酶活性。1-MCP处理抑制了果实成熟衰老,在贮藏前期抑制了AAT的活性,延迟了AAT活性高峰出现的时间,并显著降低了AAT的活性。对AheAAT序列分析发现,其全长1380 bp,编码459个氨基酸。AheAAT具有酰基转移酶的保守结构域H-x-x-x-D和L-x-x-YYPLAGR活性位点基序,D(N)F(V)GWG附加基序,属于BAHD醇酰基转移酶家族,其氨基酸序列与苹果和梨的相似性最高。同时,从菠萝蜜果实中克隆获得2个ERF转录因子,分别命名为AheERF1/2,其中AheERF1全长648 bp,编码215个氨基酸,与苹果和菜豆的同源性最高;AheERF2全长657 bp,编码218个氨基酸,与苹果、番木瓜的同源性最高。AheERF1/2的氨基酸序列含有64/65个氨基酸残基组成的AP2/ERF保守序列,具有ERF家族转录因子的特征序列。ETH处理使果实中AheAAT表达高峰提前出现,并且下调了AheAAT的表达。ETH处理对AheERF1/2转录因子的影响与AheAAT相同,并且ETH降低了AheERF1/2转录因子与AheAAT的相关性。1-MCP处理延长了菠萝蜜果实的贮藏期。在1-MCP处理后的贮藏前期,AheAAT和AheERF1/2转录因子的表达呈现显著降低。�【Objective】The effects of Ethephon(ETH)and 1-methylcyclopropene(1-MCP)on AAT activities,the expression of AheAAT gene and AheERF1/2 transcription factor were studied during ripening of jackfruit to provide a theoretical basis for further understanding the role of AATS in the synthesis of esters and its regulation.【Method】The test material was jackfruit of Haida2.During the full bloom period,the fruit with the representative flowering period was labeled and harvested about 150 days after flowering.The fruits with the same size and maturity and no disease and insect damage were selected to divide into three groups for treatment.The first group was immersed in 1000 mg·L-1 ETH solution for 2-3 min;the second group was fumigated with 0.5 mg·L-11-MCP for 15 h;the third group,as a control group,was naturally matured at 22℃and 90%relative humidity conditions.During fruit ripening,samples were taken periodically with 3-5 fruits per times.The flesh of the jackfruit was frozen in liquid nitrogen and then stored in an ultra-low temperature refrigerator at-80℃for determination of AAT activities,the expression of AheAAT gene and AheERF1/2 of jackfruit.【Result】By analyzing the activity of AAT enzyme,it was found that ETH treatment promoted the ripening and senescence of jackfruit,advanced the peak of the activity peak of AAT,but reduced AAT enzyme activity.1-MCP treatment inhibited the ripening and senescence of fruits by inhibiting the AAT activity in the pre-storage period,delaying the time of peak AAT activity,and significantly reducing the activity of AAT enzyme.Sequence analysis of AheAAT gene showed that the total length of AheAAT gene was 1380 bp,encoding 459 amino acids.AheAAT had a conserved domain H-x-x-x-D and an L-x-x-YYPLAGR active site motif,a D(N)F(V)GWG add-on motif.The AheAAT gene belonged to the BAHD alcohol acyltransferase family,and its amino acid sequence had the highest similarity to apples and pears.At the same time,two ERF transcription factors,named AheERF1/2,were cloned from the fruit

关 键 词：菠萝蜜 乙烯利 1-MCP AheAAT 

分 类 号：S66[农业科学—果树学]