薛雯心 劉立業(yè) 袁金月 劉琪 魏鐵鎖 鄭彥杰 李秀霞
摘要[目的]研究黨參毛狀根的誘導(dǎo)條件。[方法]利用發(fā)根農(nóng)桿菌A4、C58C1和A1476進行毛狀根誘導(dǎo),探討發(fā)根農(nóng)桿菌種類、外植體類型、菌液濃度、侵染時間和共培養(yǎng)天數(shù)等因素對黨參毛狀根誘導(dǎo)的影響。[結(jié)果]3種發(fā)根農(nóng)桿菌中A4誘導(dǎo)率最高;莖段的誘導(dǎo)率最高;發(fā)根農(nóng)桿菌濃度在OD600為0.4時誘導(dǎo)效果較好;較佳侵染時間為5 min;共培養(yǎng)時間為2 d時誘導(dǎo)率較高;頭孢噻肟鈉濃度為300 mg/L時除菌效果較好。[結(jié)論]該試驗為大量生產(chǎn)毛狀根以及毛狀根的后續(xù)研究提供試驗基礎(chǔ)。
關(guān)鍵詞黨參;毛狀根;誘導(dǎo)條件;發(fā)根農(nóng)桿菌
中圖分類號S567.5+3文獻標(biāo)識碼A文章編號0517-6611(2017)10-0121-03
Study on the Induction Condition of Hairy Roots of Codonopsis pilosula
XUE Wenxin,LIU Liye,YUAN Jinyue,LI Xiuxia* et al
(College of Life Science, Jiamusi University, Jiamusi,Heilongjiang 154007)
Abstract[Objective] The research aimed to study the induction condition of hairy roots of Codonopsis pilosula.[Method] The hairy roots were induced by Agrobacterium rhizogenes strains of A4, C58C1 and A1476,the effects of Agrobacterium rhizogenes species, explants types, concentration of bacterium,infection time, cocultivation period on hairy roots inductivity of Codonopsis pilosula were discussed.[Result] Among the three Agrobacterium rhizogenes,the induction rate of A4 was the highest.The induction rate of stem was the highest.The inductivity at OD600 0.4 was better performing.Better infection time was five minutes.The inductivity was higher when the time of cocultivation was two days. When the concentration of cefotaxime sodium was 300 mg/L, the sterilization effect was better.[Conclusion] This test provides experimental basis for the mass production of hairy roots and followup study of the hairy roots.
Key wordsCodonopsis pilosula;Hairy roots;Inducement condition;Agrobacterium rhizogenes
黨參[Codonopsis pilosula(Franch.)Nannf.]屬桔梗科植物[1],多年生草質(zhì)藤本,以干燥根入藥[2],為我國常用大宗藥材之一[3]。我國是黨參屬藥材的主要產(chǎn)區(qū)和資源分布中心,全世界有黨參屬植物40 多種,我國就有39 種之多[4],主產(chǎn)于山西、陜西、甘肅等地[5]。幾十年來,在原有研究基礎(chǔ)上,國內(nèi)外對黨參化學(xué)成分進行了深入研究[6]。黨參屬藥用植物所含化學(xué)成分相似,主要有聚炔類、萜及三萜皂苷類、苯丙素類、茲醇、生物堿、倍半萜內(nèi)酯、香豆素等多種類型的化學(xué)成分。從黨參中還分離得到多糖類、甾類、揮發(fā)油成分、多種氨基酸和微量元素[7]。
現(xiàn)代藥理研究表明,黨參具有調(diào)節(jié)血糖、促進造血、降壓、抗缺氧、耐疲勞、增強機體免疫力、調(diào)節(jié)胃腸收縮及抗?jié)兊榷嘀刈饔肹8],并常作為人參的替代品[9]。
發(fā)根農(nóng)桿菌(Agrobacterium rhizogenes)是一種根瘤菌科的革蘭氏陰性菌[10],其侵染植物后,會使植物產(chǎn)生大量的不定根,這些不定根不斷分支,表現(xiàn)出毛狀,稱之為毛狀根(Hairy Roots)[11]?!?br>