作　　者： ; ; ; ; ; (李振勇）;

机构地区： 西北大学生命科学学院

出　　处： 《西北大学学报（自然科学版）》 2006年第1期89-92,共4页

摘　　要： 目的用mRNA差异显示技术分离小麦盐胁迫应答cDNA,为进一步研究小麦耐盐机理奠定基础。方法将小麦(Triticum aesticum L.)耐盐品系宝丰7228r种子分别置于含NaC l 0‰和12‰的Hoagland培养液中生长,7d后分别取叶片,提取总RNA并分离出其中的mRNA,通过锚锭引物O ligo dT10GC反转录和9个10核苷酸随机引物进行PCR扩增。结果DDRT-PCR结果显示,有4个差异DNA片段只在盐胁迫的小麦耐盐品系基因组中表达,而在对照中没有出现。这4个差异cDNA片段分别命名为ts01(260 bp),ts02(330 bp),ts03(420 bp),ts04(600 bp)。4个差异cD-NA片段的RNA杂交结果显示,只有ts04存在明显差异,在盐胁迫条件下有杂交斑点而在对照中没有杂交斑点,其余3个cDNA片段在盐胁迫和对照中都没有杂交斑点。进一步将ts04克隆并进行DNA序列测定及同源性分析。结论ts04可能是耐盐相关cDNA片段,该片段核酸序列与麦属抗性相关的肌动蛋白基因有23%同源。 Aim To extract the salt responsive cDNA in wheat by mRNA differential display based on a foundation for further research the mechanism of wheat salt tolerance. Methods Total RNAs of the leaves in wheat （ Tritlcum aestivum L. Baofeng 7228r） which cultured in 0‰, 12‰ NaCl condition were isolated respectively for mRNA differential display analysis, and the reverse transcription and PCR were performed by Oligo dT10GC and nine 10-oliqonucleotide arbitrary primers. Results It showed that the differential cDNA fragments of ts01 （260bp）, ts02 （330bp）, ts03 （420bp）, ts04 （600bp）, only appeared in the leaves of wheat Baofeng 7228r cultured in 12‰ NaCl stress condition. After detecting by RNA dot hybridization, only ts04 revealed the difference between the RNAs of the leaves in wheat Baofeng 7228r cuhured in 12‰ and 0‰ NaCI condition. Then,ts04 was cloned and sequenced. Conclusion Ts04 was confirmed as the salt responsive cDNA. After searching in GeneBank,it has part homology with the Triticum monococcum actin （ACT-1） gene.

关 键 词： 小麦 差异显示 耐盐性

领　　域： [生物学]