[1]付晓佳,张倩,李琳,等.农杆菌介导烟草抗真菌β-1,3-葡聚糖酶基因转化洋桔梗[J].江苏农业学报,2016,(01):44-50.[doi:10.3969/j.issn.1000-4440.2016.01.007]
 FU Xiao-jia,ZHANG Qian,LI Lin,et al.Agrobacterium-mediated transformation of tobacco β-1,3-glucanase gene into Eustoma grandiflorum (Raf.) Shinn.[J].,2016,(01):44-50.[doi:10.3969/j.issn.1000-4440.2016.01.007]
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农杆菌介导烟草抗真菌β-1,3-葡聚糖酶基因转化洋桔梗()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2016年01期
页码:
44-50
栏目:
遗传育种·生理生化
出版日期:
2016-01-08

文章信息/Info

Title:
Agrobacterium-mediated transformation of tobacco β-1,3-glucanase gene into Eustoma grandiflorum (Raf.) Shinn.
作者:
付晓佳张倩李琳乔宁宁王转梅陈崇顺
(南京师范大学生命科学学院/江苏省生物多样性与生物技术重点实验室,江苏南京210023)
Author(s):
FU Xiao-jiaZHANG QianLI LinQIAO Ning-ningWANG Zhuan-meiCHEN Chong-shun
(College of Life Sciences, Nanjing Normal University/Jiangsu Key Laboratory of Biodiversity and Biotechnology, Nanjing 210023, China)
关键词:
洋桔梗β-13-葡聚糖酶基因农杆菌介导法灰霉病抗性
Keywords:
Eustoma grandiflorumβ-13-glucanase geneAgrobacterium-mediated transformationresistance to Botrytis cinerea
分类号:
Q789
DOI:
10.3969/j.issn.1000-4440.2016.01.007
文献标志码:
A
摘要:
为获得洋桔梗抗真菌病害新品种,本研究将烟草抗真菌β-1,3-葡聚糖酶基因(Glu)与质粒pBI121重组,构建了植物表达载体pBI121-Glu,利用农杆菌介导法转化洋桔梗品种Double Mariachi Pink叶块,共获得21个卡那霉素抗性株系。对抗性株系进行PCR扩增,获得10个阳性株系。选取其中生长良好的5个株系进行RT-PCR检测,鉴定出3个株系(EG6、EG11和EG17)为转基因正常转录株系。EG6、EG11和EG17株系及非转基因对照株系的β-1,3-葡聚糖酶的平均活力分别为0.301 U、0.243 U、0.401 U及0.236 U,其中EG6和EG17株系的平均活力显著高于非转基因对照株系。洋桔梗离体叶片的抗灰霉病鉴定显示,EG17病斑面积(0.432 cm2)显著小于非转基因对照株系(2.156 cm2)。此外,EG17对灰霉病的抗性还表现为发病延迟、病斑面积扩大速度减慢等。
Abstract:
In order to enhance the resistance to fungi diseases, an antifungal β-1,3-glucanase gene from tobacco was transferred into Double Mariachi Pink lisianthus [Eustoma grandiflorum (Raf.) Shinn.] through Agrobacterium-mediated transformation by constructing the plant expression vector of pBI121-Glu . 21 kanamycin-resistant lines were obtained, and 10 lines carrying the target gene were detected by PCR. RT-PCR analysis confirmed that the β-1,3-glucanase gene was expressed in three lines, EG6, EG11 and EG17, which showed average β-1,3-glucanase activities of 0.301 U, 0.243 U and 0.401 U, higher than that of non-transformed line (0.236 U). Compared with non-transformed plants, resistance to Botrytis cinerea in the transgenic line EG17 was improved as smaller lesion area, slower lesion expansion speed and delayed onset of B.cinerea.

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备注/Memo

备注/Memo:
收稿日期:2015-05-29基金项目:江苏高校优势学科建设工程资助项目作者简介:付晓佳(1991-),女,山东淄博人,硕士研究生,主要从事植物生物技术与分子生物学研究。(E-mail)fuxiaojia716@163.com通讯作者:陈崇顺,(E-mail)chenchongshun@njnu.edu.cn
更新日期/Last Update: 2017-05-08