【摘要】：本研究以野生型三生煙(Nicotiana tabacum cv.Xanthi nc.)及T_2、T_3代轉(zhuǎn)hpa Xm(即hpa1Xm)基因煙草和轉(zhuǎn)hpa Xm N-端缺失突變體(標(biāo)記為hpa Xm△LP)煙草為試材,對(duì)目標(biāo)基因的整合表達(dá)、葉綠素含量及防御酶活性進(jìn)行檢測(cè)。結(jié)果表明,卡那霉素抗性篩選、PCR及RT-PCR檢測(cè)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(zhuǎn)錄水平上正常表達(dá),且兩個(gè)世代中穩(wěn)定遺傳。葉綠素含量測(cè)定中,轉(zhuǎn)基因煙草含量顯著高于野生型植株,同種轉(zhuǎn)基因植株T_2代含量顯著高于T_3代,而轉(zhuǎn)hpa Xm基因煙草的總?cè)~綠素含量比轉(zhuǎn)hpa Xm△LP基因煙草的高,無(wú)顯著性差異。防御酶活性的測(cè)定,表明轉(zhuǎn)基因煙草的PAL、POD和PPO活性皆顯著高于野生型植株;而轉(zhuǎn)hpa Xm基因煙草的PAL和POD活性顯著高于轉(zhuǎn)hpa Xm△LP基因煙草;同品種轉(zhuǎn)基因煙草中,T_2代PAL活性顯著高于T_3代。說(shuō)明hpa1Xm基因在煙草中表達(dá)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(zhuǎn)基因煙草之間表現(xiàn)有所差異。初步探索了hpa Xm的功能、遺傳穩(wěn)定性及信號(hào)肽類似序列對(duì)hpa Xm的表達(dá)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(xué)熱帶農(nóng)林學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(31360029;31160359) 國(guó)家農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系建設(shè)(CARS-34-GW8) 教育部博士生基金(20124601110004;20104601110004) 國(guó)家重大基礎(chǔ)研究計(jì)劃項(xiàng)目(2011CB111612)共同資助
【分類號(hào)】：Q943.2;S572
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本文編號(hào)：2120309