不同培养基对苜蓿花药愈伤组织继代和分化的影响

将7种苜蓿品种(品系)(MT-2、MT-146、MT-328、MT-335、新牧1号、新疆大叶和KS220-97)的混合花药接种于N6大量元素+MS微量元素+MS铁盐+B5有机物+2,4-D 2mg/L+6-BA 0.5 mg/L的固液双层培养基上,诱导出愈伤组织,再转接到不同成分及添加物的培养基中培养,观察培养效果.结果表明:①在含MS、SH、N6的大量元素的培养基上的初代培养, MS或SH的效果好于N6,愈伤组织第5～9次继代培养中,N6大量元素培养基的继代培养效果最好.②花药愈伤组织继代培养基中添加AC、LH、PVP-40对愈伤组织的生长和防止褐变有明显作用.③继代培养中期,添加NAA超过0.5 mg/L时易褐化.④在N6大量元素培养基上的愈伤分化、生长表现好于其它培养基,1/2MS大量元素次之.⑤在分化培养的后期不加激素,有利于愈伤组织的生长.

Effect of Different Culture Medium on Callus Subculture and Differentiation from Alfalfa Anther

Mixed anther of seven alfalfa varieties or Strains(MT-2,MT-146,MT-328,MT-335,Xinmu No.1,Xinjiang Daye and KS220-97) were inoculated on the double layer medium which contained N6 macroelement,MS microelements,MS molysite,B5 organic compound,2,4-D 2 mg/L,6-BA 0.5 mg/L,and induced callus,then the callus was induced on the medium of the different composition and the additives,to observe its result of culture.The result indicated: ①In accordance with culturing in the medium containing MS,SH,N6 macroelement,respectively,the effect of MS or SH macroelement in the primary culture is better than that of N6 macroelement,and the effect based on N6 macroelement is the best from 5th to 9th subculture.② The subculture medium of the anther callus with AC、LH and PVP-40 added can distinctly promote the growth of callus and prevent it from browning.③ In the metaphase of subculture,if the addition of NAA is more than 0.5mg/l,it is easy to be browning.④ The growth manifestation of the callus differentiation for N6 macroelement is better compared with other mediums,then 1/2MS macroelement medium is following.⑤ The anaphase of differentiation culture without hormone benefits the growth of the callus.