中国水仙凝集素基因在大花烟草中的定量表达及抗蚜性分析

doi:10.7668/hbnxb.2017.04.013

摘要/Abstract

摘要： 为了确定中国水仙凝集素基因NTL1的抗蚜功能及培育具有抗蚜功能的大花烟草，利用qPCR方法，研究了中国水仙凝集素基因（NTL1）在转基因大花烟草4个株系中的相对表达情况；利用自然感蚜、活体接种和离体接种方法研究了不同转基因株系的抗蚜能力。结果表明，NTL1在不同株系中的相对表达量差异显著，从高到低依次为3，4，1，2；抗蚜虫试验结果显示：转基因烟草具有一定抗蚜性，但不同烟草株系的抗蚜能力有所不同；活体转基因烟草单株与离体烟草叶片的平均蚜虫密度抑制率分别为8.22%~76.61%，4.62%~65.65%；转基因大花烟草可加速蚜虫的死亡。对比NTL1在不同转基因大花烟草株系中的相对表达量及抗蚜效果，发现NTL1在不同株系中的相对表达量与植株抗蚜性具有高度一致性。

关键词: 中国水仙, 凝集素基因, qPCR, 大花烟草, 抗蚜性

Abstract: The expression of the lectin gene (NTL1) from Narcissus tazzetta var. chinensis in Nicotiana alata was studied by qPCR method in order to determine the aphid-resistant function of NTL1 and cultivate N. alata with aphid-resistant. By means of susceptible to aphid,inoculation in vivo and in vitro,the resistance to aphid in different transgenic tobacco was studied. The result showed that NTL1 gene had been integrated into the tobacco,but the expression of different tobacco was different,the expression of NTL1 in tobacco by descending order was 3,4,1,2. The bioassay of aphid indicated that transgenic tobacco had a certain resistance to aphid,but the capacity of aphid-resistance in different tobacco plant was slightly different;The average density inhibition rate of aphid in inoculation in vivo and in vitro was 8.22%-76.61% and 4.62%-65.65%. The transgenic tobacco could speed up the aphid's death. The relative expression of NTL1 gene in different transgenic tobacco and the result of resistance to aphid were compared,showed that two results came with a high degree of consistency.

Key words: Narcissus tazetta var. chinensis, Lectin gene, qPCR, Nicotiana alata, Resistance to aphid

中图分类号:

S433.1

杨会苗, 陈段芬. 中国水仙凝集素基因在大花烟草中的定量表达及抗蚜性分析[J]. 华北农学报, 2017, 32(4): 78-84. doi: 10.7668/hbnxb.2017.04.013.

YANG Huimiao, CHEN Duanfen. Analysis of qPCR and Resistance to Aphid of NTL1 Gene from Narcissus tazetta var. chinensis in Nicotiana alata[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(4): 78-84. doi: 10.7668/hbnxb.2017.04.013.

http://www.hbnxb.net/CN/Y2017/V32/I4/78

参考文献

[1] Van Damme E M,Peumans W J,Pusztal A,et al. Handbook of plant lectins:ProPertis and bio-medical application[M]. Chichester:John Willey and Sons Ltd,1997.
[2] 于敏,袁萍萍,章莹莹. 植物凝集素的部分生物学作用分析[J]. 安徽农业科学,2002,30(1):155-156.
[3] 周晓宇,陈杰,杨敬,等. 植物凝集素及其在抗虫基因工程中的应用[J]. 山地农业生物学报,2010,29(3):255-260.
[4] 邓青,陈巨莲,程登发,等. 植物凝集素及其抗蚜作用研究进展[J]. 应用昆虫学报,2013,50(1):268-275.
[5] Czapla T H,Lang B A. Effect of plant lectins on the larval develepment of European corn borer (Lepidoptera:Pyralidae) and Southern corn rootworm (Coleoptera:Chrysomelidae)[J]. J Econ Etomol,1990,83(6):2481-2485.
[6] 郭佩佩. 大豆凝集素基因lec-s 的克隆及转化烟草的研究[D]. 南京:南京农业大学,2012.
[7] 田莉莉. 转ASGNA拟南芥鉴定,抗虫性分析和对棉花的遗传转化[D]. 西安:陕西师范大学,2015.
[8] 张晓英,甘敬,尹伟伦,等. 国槐转雪花莲凝集素基因及抗蚜性[J]. 林业科学,2010,46(2):51-55,封三.
[9] Hilder V A,Powell K S,Gatehouse A,et al. Expression of snowdrop lectin in transgenic tobacco plants results in added protection against aphids[J]. Transgenic Research,1995,4(1):18-25.
[10] 吴昌银,叶志彪,李汉霞,等. 雪花莲外源凝集素基因转化番茄[J]. 植物学报,2000,42(7):719.
[11] 段晓亮,许兰杰,侯起岭,等. 转人工合成中国水仙凝集素基因(sNTL)小麦的获得及其抗麦长管蚜效果分析[J]. 华北农学报,2016,31(1):8-14.
[12] Miao J,Wu Y Q,Xu W G,et al. The impact of transgenic wheat expressing GNA (snowdrop lectin) on the aphids Sitobion avenae, Schizaphis graminum,and Rhopalosiphum padi[J]. Environmental Entomology,2011,40(3):743-748.
[13] Aasen S S,Hagvar E B. Effect of potato plants expressing snowdrop lectin (GNA) on the performance and colonization behaviour of the peach-potato aphid Myzus persicae[J]. Acta Agriculturae Scandinavica Section B-Soil and Plant Science,2012,62(4):352-361.
[14] 周英,谢红卫,刘长爱,等. 豆科凝集素基因Le4 的克隆及其表达产物对蚜虫的抗性[J]. 基因组学与应用生物学,2016(12):3474-3480.
[15] 刘玎,陈劲,刘志,等. 掌叶半夏凝集素基因PPA2 抗蚜功能分析[J]. 生物技术通报,2016,32(10):180-187.
[16] 常淑芬. 人工合成的抗蚜基因(ASGNA)在拟南芥中的表达及抗蚜性分析研究[D]. 西安:陕西师范大学,2013.
[17] 段晓亮. 三种植物凝集素基因在转基因小麦中的表达及其抗蚜效果分析[D]. 北京:中国农业大学,2013.
[18] Rao K V,Rathore K S,Hodges T K,et al. Expression of snowdrop lectin (GNA) in transgenic rice plants confers resistance to rice brown plant hopper[J]. The Plant Journal,1998,15(4):469-477.
[19] Powell K S,Gatehouse A M R,Hilder V A,et al.Antifeedent effects of plant lectins and enzyme on the adulft stage of the rice brown planthopper, Nilaparvata lugens[J]. Entom Experimet Applicta,1995,75(1):51-59.
[20] Habibi J,Backus E A,Czapla T H. Plant lectins affect survival of the potato leafhopper(Homoptera:Cicadellidae)[J].J Econ Etomol,1993,86(86):945-951.
[21] 王志斌,李学勇,郭三堆. 植物凝集素与抗虫基因工程[J]. 生物技术通报,1998(2):5-10.
[22] 邹玉,邵鸣,王涛,等. 蚜虫防治研究进展[J]. 中国林副特产,2016(6):89-91.
[23] 张海娜,钱玉源,刘祎,等. 蚜虫防治研究概况及在棉花上的应用[J]. 农学学报,2015,5(8):36-39.
[24] Bostwick D K,Skaggs M I,Thompson G A. Organization and characterization of Cucurbita phloem lectin genes[J].Plant Molecular Biology,1994,263(3):887-897.
[25] 王关林,刘彦泓,郭绍华,等. 雪花莲凝集素基因转化菊花及转基因植株的抗蚜性研究[J]. 遗传学报,2004,31(12):1434-1438.
[26] 高泽发,陈绪清,杨凤萍,等. 人工合成gna 基因在小麦中的表达及其抗蚜虫效果研究[J]. 农业生物技术学报,2006,14(4):559-564.
[37] 张小霞,曹素梅,梁振普,等. 植物凝集素在抗刺吸式害虫转基因工程中的应用[J]. 植物保护,2010,36(4):23-28.
[28] 周岩,田颖川. 转雪花莲外源凝集素基因烟草对桃蚜的抑制作用[J]. 生物工程学报,1998,14(1):13-19.
[29] 于小清,陈段芬. 中国水仙凝集素基因的克隆及性质预测[J]. 华北农学报,2009,24(2):47-50.
[30] 曹如槐,粟克恭,王晓玲. 农作物抗病虫性鉴定方法[M]. 北京:中国农业出版社,1992.
[31] 徐雪莲,张银东,符悦冠,等. 不同西瓜品种抗蚜性鉴定[J]. 中国蔬菜,2013,10(20):82-86.

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