【摘要】：本研究以野生型三生煙(Nicotiana tabacum cv.Xanthi nc.)及T_2、T_3代轉hpa Xm(即hpa1Xm)基因煙草和轉hpa Xm N-端缺失突變體(標記為hpa Xm△LP)煙草為試材,對目標基因的整合表達、葉綠素含量及防御酶活性進行檢測。結果表明,卡那霉素抗性篩選、PCR及RT-PCR檢測證明,外源目的基因hpa Xm和hpa Xm△LP整合到煙草基因組中,在轉錄水平上正常表達,且兩個世代中穩(wěn)定遺傳。葉綠素含量測定中,轉基因煙草含量顯著高于野生型植株,同種轉基因植株T_2代含量顯著高于T_3代,而轉hpa Xm基因煙草的總葉綠素含量比轉hpa Xm△LP基因煙草的高,無顯著性差異。防御酶活性的測定,表明轉基因煙草的PAL、POD和PPO活性皆顯著高于野生型植株;而轉hpa Xm基因煙草的PAL和POD活性顯著高于轉hpa Xm△LP基因煙草;同品種轉基因煙草中,T_2代PAL活性顯著高于T_3代。說明hpa1Xm基因在煙草中表達能顯著提高其葉綠素含量及抗病防御酶活性水平,且不同世代的轉基因煙草之間表現有所差異。初步探索了hpa Xm的功能、遺傳穩(wěn)定性及信號肽類似序列對hpa Xm的表達及功能的影響。
[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.
【作者單位】： 海南大學熱帶農林學院;
【基金】：國家自然科學基金(31360029;31160359) 國家農業(yè)產業(yè)技術體系建設(CARS-34-GW8) 教育部博士生基金(20124601110004;20104601110004) 國家重大基礎研究計劃項目(2011CB111612)共同資助
【分類號】：Q943.2;S572
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本文編號：2120309