条锈菌诱导下的小麦叶片总RNA提取方法的比较及LD-PCR扩增

Comparison of Different Protocols of Extracting Total RNA from Rust Induced Wheat Leaves and LD-PCR Amplification

为寻找一种既能提取到高质量高产量小麦RNA、又能提取到高质量条锈菌RNA的方法,利用Trizol法、Licl法、SDS法和Bizol法分别从条锈菌诱导的小麦叶片中提取了总RNA,并对Trizol法和Bizol法提取的RNA进行了长距离PCR（LD-PCR）扩增。结果表明,Trizol法、Licl法、SDS法和Bizol法提取的RNA产量（μg/100mg鲜重叶片）分别为4.72、1.15、1.56、10.43μg/100 mg,Trizol法和Bizol法提取的RNA产量明显高于Licl法和SDS法。Trizol法提取的RNA合成的cDNA片段大小主要集中在0.25-2.0 kb范围内,而Bizol法提取的RNA合成的cDNA片段大小主要集中在0.5-5.0 kb范围内,Bizol法提取的RNA反转录合成的cDNA质量好于Trizol法。Bizol法同样能提取到高质量的条锈菌RNA,而且提取RNA的成本较低。综合各种因素,Bizol法是条锈菌诱导下的小麦叶片总RNA提取的较理想方法。

To find a protocol of extracting RNA that has high production and good quantity from both wheat leaves and rust, Trizol, Licl,SDS and Bizol protocols were used for extracting total RNA from rust induced wheat leaves, total RNA extracted by Trizol and Bizol protocols was amplified by long distance PCR （LD-PCR）. The results showed that total RNA production of wheat leaves （μg/100 mg leaves） extracted by Trizol, Licl, SDS and Bizol protocols were 4.72,1.15,1.56,10.43 kg/100 mg, respectively, total RNA production of wheat leaves extracted by Trizol and Bizol protocols were obvi- ously higher than that by Licl and SDS protocols, cDNA synthesized from RNA extracted by Trizol was mainly 0.25-2.0 kb, cDNA synthesized from RNA extracted by Bizol was mainly 0.5-5.0 kb, cDNA synthesized from RNA extracted by Bizol was better than that by Trizol. RNA of rust can also be extracted by Bizol and it cost less. Considering all aspects, Bizol protocol is the best one to extract RNA from rust induced wheat leaves.