转hpa1Xm基因烟草后代的分子检测及苗期生理生化指标测定
发布时间：2018-07-13 18:11
【摘要】：本研究以野生型三生烟(Nicotiana tabacum cv.Xanthi nc.)及T_2、T_3代转hpa Xm(即hpa1Xm)基因烟草和转hpa Xm N-端缺失突变体(标记为hpa Xm△LP)烟草为试材,对目标基因的整合表达、叶绿素含量及防御酶活性进行检测。结果表明,卡那霉素抗性筛选、PCR及RT-PCR检测证明,外源目的基因hpa Xm和hpa Xm△LP整合到烟草基因组中,在转录水平上正常表达,且两个世代中稳定遗传。叶绿素含量测定中,转基因烟草含量显著高于野生型植株,同种转基因植株T_2代含量显著高于T_3代,而转hpa Xm基因烟草的总叶绿素含量比转hpa Xm△LP基因烟草的高,无显著性差异。防御酶活性的测定,表明转基因烟草的PAL、POD和PPO活性皆显著高于野生型植株;而转hpa Xm基因烟草的PAL和POD活性显著高于转hpa Xm△LP基因烟草;同品种转基因烟草中,T_2代PAL活性显著高于T_3代。说明hpa1Xm基因在烟草中表达能显著提高其叶绿素含量及抗病防御酶活性水平,且不同世代的转基因烟草之间表现有所差异。初步探索了hpa Xm的功能、遗传稳定性及信号肽类似序列对hpa Xm的表达及功能的影响。
[Abstract]:In this study, wild type Sansheng tobacco (Nicotiana tabacum cv. Xanthi NC.) The transgenic hpa Xm (hpa1Xm) gene and hpa Xm N- terminal deletion mutant (labeled hpa Xm LP) were used to detect the integrated expression, chlorophyll content and defense enzyme activity of the target gene. The results showed that kanamycin resistant gene hpa Xm and hpa Xm LP were integrated into the tobacco genome and expressed normally at the transcriptional level. The content of chlorophyll in transgenic tobacco was significantly higher than that in wild-type tobacco, and the content of total chlorophyll in transgenic tobacco with hpa Xm gene was higher than that of transgenic tobacco with hpa Xm LP gene. There was no significant difference. The activity of defense enzymes in transgenic tobacco was significantly higher than that in wild-type tobacco, while the activity of pal and POD in transgenic tobacco with hpa Xm gene was significantly higher than that in transgenic tobacco with hpa Xm LP gene. The pal activity in the second generation of transgenic tobacco was significantly higher than that in the third generation. The results showed that the expression of hpa1Xm gene in tobacco could significantly increase the chlorophyll content and the activity of disease resistance and defense enzymes, and the difference of the expression of hpa1Xm gene between different generations of transgenic tobacco was also found. The function of hpa Xm, the effect of genetic stability and similar sequence of signal peptide on the expression and function of hpa Xm were preliminarily investigated.
【作者单位】： 海南大学热带农林学院;
【基金】：国家自然科学基金(31360029;31160359) 国家农业产业技术体系建设(CARS-34-GW8) 教育部博士生基金(20124601110004;20104601110004) 国家重大基础研究计划项目(2011CB111612)共同资助
【分类号】：Q943.2;S572
，
本文编号：2120309
[Abstract]:In this study, wild type Sansheng tobacco (Nicotiana tabacum cv. Xanthi NC.) The transgenic hpa Xm (hpa1Xm) gene and hpa Xm N- terminal deletion mutant (labeled hpa Xm LP) were used to detect the integrated expression, chlorophyll content and defense enzyme activity of the target gene. The results showed that kanamycin resistant gene hpa Xm and hpa Xm LP were integrated into the tobacco genome and expressed normally at the transcriptional level. The content of chlorophyll in transgenic tobacco was significantly higher than that in wild-type tobacco, and the content of total chlorophyll in transgenic tobacco with hpa Xm gene was higher than that of transgenic tobacco with hpa Xm LP gene. There was no significant difference. The activity of defense enzymes in transgenic tobacco was significantly higher than that in wild-type tobacco, while the activity of pal and POD in transgenic tobacco with hpa Xm gene was significantly higher than that in transgenic tobacco with hpa Xm LP gene. The pal activity in the second generation of transgenic tobacco was significantly higher than that in the third generation. The results showed that the expression of hpa1Xm gene in tobacco could significantly increase the chlorophyll content and the activity of disease resistance and defense enzymes, and the difference of the expression of hpa1Xm gene between different generations of transgenic tobacco was also found. The function of hpa Xm, the effect of genetic stability and similar sequence of signal peptide on the expression and function of hpa Xm were preliminarily investigated.
【作者单位】： 海南大学热带农林学院;
【基金】：国家自然科学基金(31360029;31160359) 国家农业产业技术体系建设(CARS-34-GW8) 教育部博士生基金(20124601110004;20104601110004) 国家重大基础研究计划项目(2011CB111612)共同资助
【分类号】：Q943.2;S572
，
本文编号：2120309
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