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

摘要:

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7 d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。

Abstract:

Genetic transformation of Solanum photeinocarpum by Agrohacterium rhizogenes R1000 and R1601． the effects of some factors on transformation efficiency and the production of secondary metabolites by hairy root culture were studied. Hairy roots were induced from leaf and stern explants with A．rhizogenes strain R1000 and R1601 ca. 7 days after inoculation Hairy root frequencies obtained from leaf and stem explants inoculated by R1000 harboring the agropine-type Ri plasmid pRiA₄b were 90.2％ and 50.0％ , respectively, while those by R1601 with the agropine-type Ri plasmid pRi1500 and pTVK291 were 92.1％ and 49.2％, respectively． Opine analysis indicated that T-DNA genes of A. rhizogenes strain R1000 and R1601 were expressed in S．photeinoccapum hairy roots. Hairy root-inducing frequency of leaf explants from S．photeinocarpum grown on MS medium supplemented with PP₃₃₃ was remarkably increased， and that of leaf explants inoculated by doubled diluting agrobacterium liquid on MS liquid medium was also increased． Hairy roots grew rapidly and the dry weight increased by 420 times after culture for 4 weeks， and the dry weight contents of total glycoalkloids and saponins were 3l and 107 times higher than those of the original ones，respectively．