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当前位置: 首页 > 文章 > 钩藤毛状根的诱导及其钩藤碱含量的测定 北方园艺 2019 (15) 49-54
Position: Home > Articles > 钩藤毛状根的诱导及其钩藤碱含量的测定 Northern Horticulture 2019 (15) 49-54

钩藤毛状根的诱导及其钩藤碱含量的测定

作  者：
吴顺;孙建春;周凯;王华英;李婷;刘姣
单  位：
中南林业科技大学生命科学与技术学院
关键词：
钩藤;AS;发根农杆菌;毛状根;钩藤碱
摘  要：
为建立钩藤毛状根的培养体系,以钩藤组培无菌苗为试材,利用发根农杆菌A4、ATCC15834、R16011侵染钩藤外植体,探讨了不同钩藤外植体、菌液浓度、AS浓度、干燥处理时间对毛状根诱导率的影响.结果表明:以钩藤叶片作为外植体,预培养3 d,以OD600=0.6的A4菌株浸泡30 min后,干燥处理1 h,转入含100μmol·L-1乙酰丁香酮无激素WPM培养基中共培养3 d,在含600 mg·L-1头孢克肟筛选培养基上能获得较高的毛状根诱导率,毛状根诱导率达53.66%.PCR扩增鉴定表明,钩藤细胞核基因组中已成功整合发根农杆菌的发根基因.钩藤毛状根在无激素1/2MS液体培养基中培养20 d后,部分毛状根中钩藤碱含量明显提高,最高含量是带钩枝条中的2.06倍,达1.211 mg·g-1.
作  者：
WU Shun;SUN Jianchun;ZHOU Kai;WANG Huaying;LI Ting;LIU Jiao;College of Life Science and Technology,Central South University of Forestry and Technology;
单  位：
WU Shun%SUN Jianchun%ZHOU Kai%WANG Huaying%LI Ting%LIU Jiao%College of Life Science and Technology,Central South University of Forestry and Technology
关键词：
Uncaria rhynchophylla;;AS;;Agrobacterium rhizogenes;;hairy root;;rhynchophylline
摘  要：
In order to found the culture system of hairy root for Uncaria rhynchophylla,the effect of different explant,concentration of bacterium,AS concentration,drying time on induction of hairy root was studied by using the sterile seedlings as a material and making Agrobacterium rhizogenes(A4,ATCC15834,R16011)infect these explants.The results showed that the induction rate of hairy root was higher when sterile leaves were used as the explant,preincubated for 3 days,soaked for 30 minutes by OD600 = 0.6 A4 solution,dried for 1 hour,cultured in hormone-free WPM medium with100μmol·L~(-1) AS,selected on screening medium with 600 mg·L~(-1) cefminox.The induction rate of hairy root was 53.66%.It was confirmed by PCR analysis that the rooting gene in Agrobacterium rhizogenes was integrated into the nuclear genome for Uncaria rhynchophylla.The content of rhynchophylline increased in some hairy roots after hairy roots were cultured in hormone-free 1/2 MS liquid medium for 20 days,in which the highest content of rhynchophylline reached a level of1.211 mg·g-1 DW and was 2.06 times as much as hook branch.

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