Development of highly regenerable callus lines and Agrobacterium-mediated transformation of Chinese lawngrass (Zoysia sinica Hance) with a cold inducible transcription factor, CBF1
Development of highly regenerable callus lines and Agrobacterium-mediated transformation of Chinese lawngrass (Zoysia sinica Hance) with a cold inducible transcription factor, CBF1
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利用冷诱导转录因子 CBF1 开发高度可再生的愈伤组织系以及农杆菌介导的中国草坪草 (Zoysia sinica Hance) 转化
DOI：
10.1007/s11240-006-9080-8
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发表时间：
2006
影响因子：
3
通讯作者：
Zhen
中科院分区：
文献类型：
--
作者：
Ruifen Li;Jianhua Wei;Hong Wang;Jie He;Zhen
中华结缕草（Zoysia sinicaHance）是一种暖季型草坪草，广泛应用于高尔夫球场、运动场和家庭草坪。为利用生物技术改良草坪草，建立了草坪草高效再生体系。通过多次继代培养，在培养基中添加6-苄基氨基嘌呤（6-BA）和调节2，4-二氯苯氧乙酸（2，4-D）浓度，获得了5个高再生率的愈伤组织系，其中愈伤组织系Zh 44用于农杆菌介导的转化。将花椰菜花叶病毒（CaMV）35 S启动子驱动的拟南芥C-repeat/dehydration-responsive elements binding factor 1（CBF 1/DREB 1b）基因导入中国草坪草中，用草铵膦筛选。采用三步选择/再生法成功获得了三个独立的转基因株系。对转基因植株进行了PCR、Southern杂交和RT-PCR检测。两个转基因株系表现出生长迟缓和分蘖减少。转基因植株在低温条件下的离子渗漏率、叶绿素含量和黄化率均显著高于野生型植株。
Chinese lawngrass (Zoysia sinicaHance) is a warm-season turfgrass, which is widely used for golf courses, athletic fields and home lawns. In order to improve Chinese lawngrass with biotechnology, an efficient plant regeneration system was established. Five highly regenerable callus lines have been developed by addition of 6-benzylaminopurine (6-BA) and adjusting concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) in culture medium after multiple subculture, of which callus line Zh44 was used forAgrobacterium-mediated transformation. The gene, encoding C-repeat/dehydration-responsive element binding factor 1 (CBF1/DREB1b) fromArabidopsisdriven by cauliflower mosaic virus (CaMV) 35S promoter, was introduced into Chinese lawngrass under selection using thebargene with glufosinate. Three independent transgenic lines were successfully obtained using a three-step selection/regeneration procedure. The transgenic plants were confirmed by polymerase chain reaction (PCR), southern blot and reverse transcriptase polymerase chain reaction (RT-PCR) analyses. Two transgenic lines exhibited growth retardation and decreased tillering. The transgenic lines have a significantly stronger chilling tolerance than that of wild type plants, measured by ion leakage, total chlorophyll content and yellowed leaf rate in chilling conditions.