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Effects of tissue culture condition on root growth and saikosaponins production in Bupleurum kaoi Liu, Chao et Chuang.
|關鍵字:||Bupleurum kaoi Liu|
Chao et Chuang
高氏柴胡之增殖，以 MS 基礎鹽類培養基，添加 0.05 mg/L NAA、0.2 mg/L BA 為增殖培養基。不定根之誘導，以B5添加0.5 mg/L IBA 為培養基。莖段倒插則以1/4 MS 添加0.1 mg/L NAA及0.5 mg/L BA 為培養基。
植株生根根系液態培養，IBA處理可使鮮重增加倍率明顯增加，SS-a、SS-d 產量分別以 9 mg/L IBA 及 7 mg/L IBA較高，SS-a + SS-d 總產量則以 7 mg/L IBA 最高，達479.2 μg/g。不同濃度蔗糖處理，結果顯示鮮重增加的倍率、乾重、SS-a、SS-d及SS-a + SS-d 總產量，均以8 % 蔗糖濃度為最高。光照培養可得到較高的鮮重增倍率、乾重及柴胡皂素的生產。繼代初始根重則以 100 ml 培養液中加入根重 1g 最適當。初代培養根及繼代初始根之根乾重均隨著培養時間增加而增加。繼代初始根重以 1 g 及 3 g 處理時，SS-a及SS-d 產量，分別以第42 天及第 35 天為最高。初代培養根 SS-a 及 SS-d產量則分別以培養第 42 天及第 21 天產量最高。
莖生根根系，在 IBA處理培養下，可使鮮重增加倍率明顯增加。乾重、SS-a 產量以 9 mg/L IBA較高，SS-d 產量則以3 mg/L IBA 較高，SS-a +SS-d總產量則以 9 mg/L IBA 最高，達841.2 μg/g。不同濃度蔗糖處理，鮮重增加的倍率及乾重，以8 % 蔗糖濃度為最高， SS-a 生產以3、5及7 % 蔗糖濃度可得到較高產量；SS-d 生產則以 1 % 蔗糖濃度可得到較高產量。光照處理下可得到較高的鮮重增倍率、乾重及柴胡皂素的生產。初代培養根鮮重增加的倍率及乾重，均隨著培養時間增加而增加，但 SS-a及 SS-d 生產，則分別以培養第 42 天及第 35 天產量最高。
SS-a、SS-d 含量 (每克鮮重) 在高氏柴胡不同來源及部位，均以實生苗根為最高；組織培養苗根最低。比較市售品、實生苗根、組織培養苗根、植株生根根系及莖生根根系之乾品柴胡皂素SS-a 、SS-d含量，柴胡皂素SS-a 含量以市售品為最高；組織培養苗根最低。柴胡皂素SS-d 含量則以莖生根根系最高；市售品最低。|
Abstract The purpose of this study was to establish tissue culture condition on the root growth and saikosaponins production in Bupleurum kaoi Liu, Chao et Chuang. MS medium containing 0.2 mg/L BA and 0.05 mg/L NAA was used as shoots' proliferation medium. The adventitious root induction of shoots was cultured on B5 medium containing 0.5 mg/L IBA. The induction of adventitious root by inverted stem segments was cultured on 1/4 MS medium containing 0.1 mg/L NAA and 0.5 mg/L BA. The roots, which were induced by shoots, increased ratio of root fresh weight by IBA treatment. Experimental data showed that higher production of saikosaponin -a (SS-a) and saikosaponin -d (SS-a) were grown at the 9 mg/L IBA and 7 mg/L IBA, respectively. Root was cultivated with 7 mg/L IBA, resulting in the highest concentration of saikosaponin-a and -d ( 479 mg/g). The ratio of maximal root fresh weight increase , dry weight, production of saikosaponin -a and saikosaponin -d was attained in B5 medium containing 8 mg/L sucrose. Initial weight of root explants subculture was 1g (DW / 100ml) better than 3g (DW / 100ml) in the liquid medium. The root explants initial weight was accumulated with time course, and higher amounts of saikosaponin-a and saikosaponin-d was attained at 42 days and 21 days. The initial weight of root explants subculture were accumulated with time course, higher amounts of saikosaponin-a and saikosaponin-d were attained at 42 days and 35 days. The roots, which were induced by stem segments, increased ratio of root fresh weight by IBA treatment. Experimental data showed that higher dried weight and production of saikosaponin-a were attained in B5 medium containing 9 mg/L IBA. The higher production of saikosaponin -d was cultured in B5 medium containing 3 mg/L IBA. The saikosaponin -a and -d production, with levels reaching 841.2 mg/g, was the most attained in B5 medium containing 9 mg/L IBA. The ratio of maximal root fresh weight increase, dry weight, production of saikosaponin -a and saikosaponin -d was attained in B5 medium containing 8 mg/L sucrose. The higher production of saikosaponin-a was attained in B5 medium containing 3, 5 and 7 mg/L sucrose. The higher production of saikosaponin-d was attained in B5 medium containing 1 mg/L sucrose. The initial weight of root explants was accumulated with time course and higher amounts of saikosaponins were attained at 42 days and 35 days. Shoots and stem segments induced roots were cultured with light was attained more higher increasing ratio of fresh weight, dry weight and production of saikosaponins than dark. In fresh weight analysis of HPLC, the highest contents of saikosaponin -a and -d were obtained on root of seedling. In dry weight analysis of HPLC, the highest contents of saikosaponins -a was obtained on market product of Radix Bupleuri. The highest contents of saikosaponin -d was obtained on stem segments induced roots and the lowest on market product of Radix Bupleuri.
|Appears in Collections:||生命科學系所|
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