红麻叶绿体DNA非编码区扩增及PCR-RFLP多态性分析  

The non-coding regions sequence characteristics of chloroplast DNA in kenaf and analysis on PCR-RFLP polymorphism

作　　者：徐建堂[1] 祁建民[1] 林培清[1] 张高阳[1] 陶爱芬[1] 方平平[1] 

机构地区：[1]福建农林大学作物遗传育种与综合利用教育部重点实验室/福建农林大学农业部东南黄红麻科学观测实验站/福建农林大学能源植物研究中心,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2013年第2期166-170,共5页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：农业部东南黄/红麻科学观测实验站项目(农科教2011-9);福建省自然科学基金资助项目(2011J05046)

摘　　要：为研究红麻叶片叶绿体DNA(cpDNA)非编码区的序列特征,采用改良CTAB方法提取红麻总DNA,以8份红麻光钝感突变体及其4份近缘种基因组DNA为模板,应用8对植物cpDNA通用引物对红麻cpDNA非编码区序列进行PCR扩增,优化cpDNA非编码序列PCR的扩增条件,应用4种限制性内切酶(EcoRⅠ、HindⅢ、TaqⅠ、AulⅠ)酶切扩增片段.结果表明:所筛选的通用引物适用于红麻cpDNA非编码序列扩增,扩增产物经1.5%琼脂糖电泳检测,片段大小为400-2000 bp,共扩增cpDNA片段大小约13000 bp.将PCR扩增产物进行酶切,每个片段均有相同的酶切位点,只有cp4引物对cpDNA的扩增产物经AluⅠ酶切获得多态性片段,红麻光钝感突变体及其对照的cpDNA存在部分变异,该结果可为进一步探讨红麻叶绿体基因组种内的变异情况提供理论依据.To study the chloroplast DNA （cpDNA） non-coding regions sequence characteristics in kenaf, with the genomic DNA of eight kenaf photoperiod-insensitive mutants and their four related species as template, we amplified cpDNA non-coding regions in kenaf using eight cpDNA universal primer pairs. Firstly, we optimized the PCR amplifing conditions of cpDNA non-coding se- quence, then digested the products by four restriction enzymes （ EcoR I , Hindm, Taq I , Au/I ）. The results showed that the u- niversal primers were suitable for the kenof cpDNA non-ceding sequence ampllilng, the products were detected by 1.5% agarese gel electrophoresis, which showed fragment size ranged from 400 to 2000 hp, the total fragments size was about 13000 bp. The digested PCR amplification products had the same digesting site, but only the product amplified by primers cp4 had polymorphie fragments af- ter being digested by Alu I , which suggested the cpDNA sequence between kenaf photoperied-insensitive mutants and controned va- rieties existed variation. Therefore, the above results could provide a theoretical basis on further exploring the relationship of chloro- plast genome variation between intraspecific kenaf.

关 键 词：红麻 叶绿体DNA PCR-RFLP 光钝感 

分 类 号：S563.5[农业科学—作物学]