CAJ | 학술논문

以湿地松(Pinus elliottii,PEE),包括古巴加勒比松(P.caribaea var.caribaea,PCC)、洪都拉斯加勒比松(P.caribaea var.hondurensis,PCH)、巴哈马加勒比松(P.caribaea var.bahmaensis,PCB)3个变种在内的加勒比松,侧枝顶芽为试验材料,使用RETSCH MM400 混合球磨仪破碎植物组织,然后利用改良CTAB法提取基因组DNA,完成48个样品仅用1 h,1个工作日可提取300个样品以上,DNA分子量大于20 kb,0.3 g样品可提取DNA 20 -60 μg,能够满足几百次PCR反应;利用所提DNA进行SSR分析,条带清晰,多态性好,说明该方法提取的DNA完全可以满足SSR反应的需要.本研究为SSR标记用于湿地松、加勒比松分子标记辅助选择育种提供了经济、高效、可靠的DNA提取方法.
이습지송(Pinus elliottii,PEE),포괄고파가륵비송(P.caribaea var.caribaea,PCC)、홍도랍사가륵비송(P.caribaea var.hondurensis,PCH)、파합마가륵비송(P.caribaea var.bahmaensis,PCB)3개변충재내적가륵비송,측지정아위시험재료,사용RETSCH MM400 혼합구마의파쇄식물조직,연후이용개량CTAB법제취기인조DNA,완성48개양품부용1 h,1개공작일가제취300개양품이상,DNA분자량대우20 kb,0.3 g양품가제취DNA 20 -60 μg,능구만족궤백차PCR반응;이용소제DNA진행SSR분석,조대청석,다태성호,설명해방법제취적DNA완전가이만족SSR반응적수요.본연구위SSR표기용우습지송、가륵비송분자표기보조선택육충제공료경제、고효、가고적DNA제취방법.
One improved CTAB method to extracted genomic DNA from terminal buds on lateral branch of slash pine and Caribbean pine was provided.According to the improved method and the use of RETSCH MM400 fragmented plant tissue for extracted of DNA,the efficiency was raised compared to the normal process.In the experiment,48 samples could be treated one time and the process could be finished in one hour,and DNA of 300 samples could be obtained in one workday.It was estimated that molecular weight of the extracted DNA is larger than 20 kb.And 20 -60 g DNA could be extracted from 0.3 g terminal bud,which would be enough for hundreds of times of PCR reaction.The extracted DNA was using in SSR analysis,the banding patterns was clear and with good polymorphism,and the quality of DNA extracted using this method was suitable for SSR analysis.This study provide an economic,efficient,and reliable DNA extraction method for SSR markers use for molecular marker-assisted selection breeding of Slash pine,Caribbean pine.