少花龙葵毛状根的诱导和次生代谢物的产生

Induction of Solanum photeinocarpum Hairy Roots and Production of Secondary Metabolites

研究了发根农杆菌(Agrobacteriumrhizogenes)对少花龙葵(Solanumphoteinocarpum)的转化和毛状根的诱导,分析了影响转化的因素,并初步检测了毛状根的生长和次生代谢物的产生。发根农杆菌菌株R1000、R1601分别感染少花龙葵的叶片、茎段,约7d后得到毛状根。菌株R1000感染的外植体的生根率分别为90.2%和50.0%,根诱导频率分别为6.2和4.6;菌株R1601感染的外植体的生根率分别为92.1%和49.2%,根诱导频率分别为6和5.5;对照两周内不出根。冠瘿碱检测证实所得毛状根为转化根。感染在MS+PP333培养基上生长的矮壮苗叶片,毛状根诱导频率显著提高;用MS液体培养基稀释1倍的菌液感染叶片,转化效率也得到提高。毛状根生长速度快,培养4周后干重增加420倍,总糖苷生物碱和皂甙含量分别为原植株根的31和107倍。

Genetictransformationof Solanum photeinocarpum by Agrobacterium rhizogenes R1000and R1601,the effectsof some factorson transformationefficiencyand the productionof secondary metabolitesby hairyrootculturewerestudied.Hairyrootswereinducedfromleafandstemexplants withA.rhizogenes strainR1000and R1601ca.7days afterinoculation.Hairyroot frequencies obtainedfrom leafand stemexplantsinoculatedby R1000harboringthe agropinetypeRi plasmid pRiA 4 b were90.2%and50.0%,respectively,whilethoseby R1601withtheagropine-typeRiplasmid pRi1500andpTVK291were92.1%and49.2%,respectively.OpineanalysisindicatedthatT-DNA genesof A.rhizogenes strainR1000andR1601wereexpressedinS.photeinocarpum hairyroots.Hairy root-inducingfrequencyof leafexplantsfromS.photeinocarpum grownon MS mediumsupplemented with PP 333 was remarkablyincreased,and that of leaf explantsinoculatedby doubleddiluting agrobacterium liquidon MS liquidmediumwas alsoincreased.Hairyrootsgrewrapidlyandthedry weightincreasedby420timesafterculturefor4weeks,and the dry weightcontentsof total glycoalkloidsandsaponinswere31and107timeshigherthanthoseof theoriginalones,respectively.