为了建立二穗短柄草高效再生体系,以二倍体二穗短柄草ABR4、ABR6和六倍体二穗短柄草ABR100、ABR101、ABR102的幼胚为外植体,研究了糖类、染色体组倍性和激素对愈伤组织诱导、分化及生根的影响。结果表明,二穗短柄草的高效再生体系的建立,其愈伤组织诱导培养基:LS+2,4-D 2.5 mg·L(-1)+麦芽糖30 g·L(-1)+琼脂6.5 g·L(-1);愈伤组织继代培养基:LS+2,4-D 1.0 mg·L(-1)+6-BA 0.2 mg·L(-1)+麦芽糖30 g·L(-1)+琼脂6.5 g·L(-1);愈伤组织分化培养基:LS+KT 0.2 mg·L(-1)+6-BA 0.5 mg·L(-1)+CuSO4 0.6 mg·L(-1)+麦芽糖30 g·L(-1)+琼脂6.5 g·L(-1);愈伤组织生根培养基:1/2 LS+IAA 0.6 mg·L(-1)+麦芽糖20 g·L(-1)+琼脂7.0 g·L(-1)。优化了二穗短柄草愈伤组织培养条件,在含有30 g·L(-1)麦芽糖的培养基上,愈伤组织出愈率最高可达96.67%,在含有0.2 mg·L(-1)KT的培养基上,分化率最高为71.25%。并进一步探讨了影响短柄草再生的主要因素。
The effects of carbon source,ecotype,hormone on induction of callus and improvement of plantlet regeneration were investigated.Immature embryos of Brachypodium distachyon from diploid accessions ABR6 and hexaploid accessions ABR102 were used as explants.The result showed that the media for callus induction was LS+2,4-D 2.5 mg·L(-1) +maltose 30 g·L(-1) +agar 6.5 g·L~(-1);the media for callus subculture was LS+2,4-D 1.0 mg·L(-1) +6-BA mg·L(-1)+ maltose 30 g·L(-1) + agar 6.5 g·L(-1);the differentiation media was LS + KT 0.2 mg·L(-1)+ CuSIO4 0.6 mg·L(-1) + maltose 30 g·L(-1)+ agar 6.5 g·L(-1);the rooting media was 1/2 LS+IAA 0.6 mg·L(-1) + maltose 20 g·L(-1) + agar 7.0 g·L(-1).The conditions of tissue culture of Brachypodium diatachyon was optimized and the induction frequency of primary calli was 96.67%on the basic media including 30 g·L(-1) maltose.The regenerated rate was 71.25%on the basic media including 0.2 g·L(-1) KT.The main factors were investigated for regeneration of Brachypodium diatachyon.