电场处理柠条种子对干旱条件下幼叶基因表达的影响  

Expression Difference of Genes in the Caragana korshinskii Young Leaves under Drought Stress by Electric Field Treatment on Seeds

作　　者：刘翔宇[1] 杨体强[1] 胡燕飞[1] 王郑[2] 哈斯阿古拉[2] 刘瑶[1] 王文星[1] 赵清春[1] 

机构地区：[1]内蒙古大学物理科学与技术学院,呼和浩特010021 [2]内蒙古大学生命科学学院,呼和浩特010021

出　　处：《西北植物学报》2014年第12期2425-2431,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(51067005)

摘　　要：以沙生植物柠条锦鸡儿为材料,设计自然对照（CK）、干旱处理对照（SCK）、电场处理（电场强度2 kV/cm,电场频率15 kHz,处理时间5 min）种子后在干旱条件下进行种子发芽（ET）,并采用3个锚定引物和26个随机引物形成78组引物,利用DDRT-PCR技术,分析电场处理柠条种子在干旱胁迫条件下生长的幼苗幼叶基因表达水平的变化,从基因转录水平探讨电场对柠条锦鸡儿抗旱性的影响机理.结果显示：（1）干旱胁迫组相对于自然对照组增加条带282条,缺失条带179条;电场处理组相对于自然对照组增加条带303条,缺失条带200条;电场处理组相对于干旱胁迫组增加条带236条,缺失条带257条.（2）回收差异片段获得电场处理诱导的3个差异表达序列（EST-1、EST 2、EST-3）,经Blastn比对,EST-1与大豆LOC100305895 （LOC100305895）一致性为76％,EST-2与鹰嘴豆UDP N-乙酰氨基葡糖肽N-乙酰葡糖胺基转移酶SEC like（LOC101506182）选择性剪切体X2一致性为88％,EST-3与百脉根克隆JCVI-FLLj-5 M12一致性为88％.（3）经Blastx比对,EST-1与1-脱氧-D-木酮糖-5-磷酸还原异构酶（390 aa）145～195氨基酸序列一致性达到52％,EST-2（308 bp）与氧连N-乙酰葡萄糖胺转移酶（744aa）706～744氨基酸序列一致性高达97％,EST-3与酰基辅酶A结合蛋白（90 aa）氨基酸序列一致性高达97％.研究结果为进一步从基因表达的角度分析电场的生物效应及其提高抗逆性的分子机理,为电场处理植物种子的农业技术应用提供理论依据.In this study,a kind of psammophytes, Caragana korshinskii seeds, were used as experiment ma- terial. The seeds were treated by electric field and the seedling growth was under drought stress conditions abd the expression difference of genes in the C. korshinshii young leaves were tested by mRNA differential display technology（DDRT-PCR）. In order to compare the gene expression difference, young leaf tissues were classified three samples include nature control（called CK）,drought stress control（called SCK） and e- lectric field treatment group（strength 2 kV/cm, frequency 15 kHz, time 5 min, called ET）. The 3 anchor primers and 26 random primers were used to form 78 primer pairs. The DDRT-PCR was carried out 234 PCR reactions for the three kinds of samples. The eDNA bands showed significant difference between three young leaf tissue samples. （1）Compared with CK, SCK appeared 282 novel bands and disappeared 179 bands, and ET appeared 303 novel bands and disappeared 200 bands. Besides, compared with SCK, ET ap- peared 236 bands and disappeared 257 bands. （2）The novel bands were recovered and three differentially expressed cDNA fragments were obtained. All of the expressed sequence tags（EST） had the homology with known cDNA clones in Genbank databases. EST-1 was similar to Glycine max uncharacterized LOC100305895（LOC100305895） and the identity was 76% ;EST-2 was similar to Cicer arietinum probable UDP-N-acetylglucosamine peptide N-acetylglucosaminyltransferase SEC-like （ LOC10t506182 ）, tran- script variant X2 and the identity was 88%;EST-3 was similar to Lotus japonicus clone JCVI-FLLj-5M12 and the identity was 88%. （3）After Blastx comparision,EST-1 was similar to the 145--195 aa of 1-deoxy- D-xylulose 5-phosphate reductoisomerase（390 aa） and the identity was 52%;EST-2 was similar to the 706 --744 aa of O-linked GlcNac transferase like protein（744 aa） and the identity was 97% jEST-3 was similar to acyl-CoA-binding protein（90 aa） and the identity was 97%. Our result pr

关 键 词：电场 柠条种子 干旱胁迫 DDRT-PCR技术 

分 类 号：Q789[生物学—分子生物学]