鸭梨POD基因的原核表达及成熟度和降温方法对其表达量的影响  

Prokaryotic Expression of POD Gene from 'Yali' Pears and Effects of Maturity and Cooling Rate on the Expression of POD Gene during Storage

作　　者：韩艳文 廉双秋 韩云云[1] 池明[1] 闫师杰[3] HAN Yanwen;LIAN Shuangqiu;HAN Yunyun;CHI Ming;YAN Shijie(College of Horticulture and Landscape, Tianjin Agricultural University, Tianjin 300384, China;Beijing Chia Tai Fruit Industry Company Limited, Beijing 101206, China;Tianjin Engineering and Technology Research Center of Agricultural Products Processing,College of Food Science and Biological Engineering, Tianjin Agricultural University, Tianjin 300384, China)

机构地区：[1]天津农学院园艺园林学院,天津300384 [2]北京正大果业有限公司,北京101206 [3]天津农学院食品科学与生物工程学院,天津市农副产品深加工技术工程中心,天津300384

出　　处：《食品科学》2017年第9期40-45,共6页Food Science

基　　金：国家自然科学基金面上项目(31471630)

摘　　要：利用半定量逆转录多聚合酶链式反应(reverse transcription polymerase chain reaction,RT-PCR)技术研究缓慢降温和急速降温两种降温方法及早采、中采、晚采3种采收成熟度鸭梨的采后果心和果肉中过氧化物酶(peroxidase,POD)基因的表达与鸭梨褐变的关系。结果发现,缓慢降温处理的鸭梨果心POD基因表达量比相同时期采摘的急速降温处理的基因表达量高,这与其对应的褐变指数相一致,说明POD与鸭梨果实褐变密切相关,缓慢降温能有效抑制鸭梨褐变。同时中采鸭梨的POD基因表达量最高,早采次之,晚采最低,这进一步说明成熟度是影响鸭梨褐变的因素之一。实验进一步构建了POD基因c DNA原核表达载体pET-32a-pbPOD,并成功诱导其在大肠杆菌BL21(DE3)中表达,为进一步研究POD表达量对鸭梨果实褐变的机理提供了参考。In the present study,the relationships between browning development and peroxidase(POD)gene expression in both core and flesh tissues of‘Yali’pears subjected to different cooling methods(rapid cooling or slow cooling),as well as those collected at different harvest dates(early,mid and late)were investigated using semi-quantitative reverse transcription polymerase chain reaction(RT-PCR).The results showed that the expression level of POD gene in pear corewith slow cooling treatment was higher than that in rapidly cooled fruits at the same harvest time,indicating that POD gene was closely related to the browning of pear core.Additionally,the POD activity was the highest in mid harvested pears,followed by early harvested pears,and late harvested pears had the lowest POD activity,which suggested that maturity played an important role in‘Yali’pear browning.Also,the prokaryotic expression of POD gene(cDNA)in‘Yali’pears was investigated.A prokaryotic expression vector(pET-32a-pbPOD)for POD gene cDNA was established,and then induced successfully to be expressed in E.coli BL21(DE3).This study may provide a foundation for studying the mechanism of the browning of‘Yali’pears.

关 键 词：鸭梨 褐变 POD基因 原核表达 成熟度 降温速率 

分 类 号：TS255.3[轻工技术与工程—农产品加工及贮藏工程]